(+)-morphinans as antagonists of toll-like receptor 9 and therapeutic uses thereof

ABSTRACT

The present invention provides (+)-morphinans comprising Toll-like receptor 9 (TLR9) antagonist activity, as well as methods for using the (+)-morphinans to treat a variety of disorders.

CROSS REFERENCE TO RELATED APPLICATIONS

This application is a continuation-in-part of U.S. patent applicationSer. No. 12/975,407, filed Dec. 22, 2010, which is acontinuation-in-part of U.S. patent application Ser. No. 12/837,545,Jul. 16, 2010, which claims the benefit of U.S. Provisional ApplicationNo. 61/286,877, filed Dec. 16, 2009 and U.S. Provisional Application No.61/226,015, filed Jul. 16, 20069, the disclosure of each is herebyincorporated by reference in its entirety.

FIELD OF THE INVENTION

The present invention generally relates to compounds and methods fortreating pain, inflammation, and other disorders. In particular, theinvention relates to (+)-morphinan compounds comprising Toll-likereceptor 9 (TLR9) antagonist activity and methods of using the compoundsto treat conditions associated with pain and inflammation.

BACKGROUND OF THE INVENTION

Activated glial cells contribute to the development and maintenance ofseveral disease states. Of particular interest is the negative impact ofactivated glial cells in the areas of chronic and acute pain,inflammatory disorders, autoimmune disorders, neurodegenerativedisorders, and cancer. Glial cells have been shown to express numerousToll-like receptors (TLRs), which are a family of highly conservedtransmembrane proteins of high functional importance in the innateimmune system. TLRs are activated by pathogen-associated molecularpatterns (PAMPs) such as lipopolysaccharide (LPS) from bacterial cellwalls, unmethylated CpG-containing DNA of viruses, and a wide variety ofadditional microbial components. Activation of TLRs in the centralnervous system is known to initiate protective pro-inflammatorysignaling cascades as part of the first line of defense against invadingpathogens. Additionally, it has been reported that chronicadministration of morphine or other opioid-receptor agonists activatesglial cells, causing the release of pro-inflammatory factors thatcounter the pain-relieving effects of the opioid. Activated glial cellshave also been shown to play a role in driving chronic pain states suchas neuropathic pain. Given these newly identified roles for glial cellsin pain, there is a need for the development of clinically useful agentsthat target glial cell activation as a means of pain control.

SUMMARY OF THE INVENTION

The present invention provides (+)-morphinan compounds that inhibit theactivation of Toll-like receptor 9 (TLR9), and consequently block glialcell activation. The compounds of the invention, therefore, may be usedto treat conditions such as traumatic pain, neuropathic pain,inflammatory disorders, acetaminophen toxicity, autoimmune disorders,neurodegenerative disorders, cancer, and inflammatory hepatic disorders.

One aspect of the present disclosure provides a method for treating ahepatic disorder in a subject in need thereof. The method comprisesadministering to the subject a compound of Formula (IIa) or apharmaceutically acceptable salt thereof:

wherein:

-   -   R is hydrogen, alkyl, alkenyl, allyl, cycloalkylmethyl,        alkylaryl, acyl, acylalkyl, acylcycloalkyl, acylaryl, acyloxy,        acyloxyalkyl, acyloxyaryl, alkoxy, or alkoxyalkyl;    -   R¹ is hydrogen, halo, hydroxy, alkoxy, alkoxyalkyl,        alkoxyalkenyl, aryloxy, alkyl, alkenyl, aryl, heteroaryl, or        amine;    -   R³ is hydroxy, alkoxy, aryloxy, acyl, acyloxy, or protected        hydroxy;    -   A₁ is hydrogen, hydroxy, keto, alkoxy, acyl, acyloxy, amino,        amide, alkylamine, hydroxyalkylamine, carboxylalkylamine,        alkylcarboxylalkylamine, arylamine, alkyl, alkenyl, aryl,        substituted alkyl, substituted alkenyl, or substituted aryl;    -   Y is hydrogen, hydroxy, alkoxy, or protected hydroxy; and    -   the dashed line represents an optional double bond.

A further aspect of the present invention encompasses a method forinhibiting ischemic-reperfusion injury in a subject in need thereof. Themethod comprises administering to the subject a compound of Formula(IIa) or a pharmaceutically acceptable salt thereof:

wherein:

-   -   R is hydrogen, alkyl, alkenyl, allyl, cycloalkylmethyl,        alkylaryl, acyl, acylalkyl, acylcycloalkyl, acylaryl, acyloxy,        acyloxyalkyl, acyloxyaryl, alkoxy, or alkoxyalkyl;    -   R¹ is hydrogen, halo, hydroxy, alkoxy, alkoxyalkyl,        alkoxyalkenyl, aryloxy, alkyl, alkenyl, aryl, heteroaryl, or        amine;    -   R³ is hydroxy, alkoxy, aryloxy, acyl, acyloxy, or protected        hydroxy;    -   A₁ is hydrogen, hydroxy, keto, alkoxy, acyl, acyloxy, amino,        amide, alkylamine, hydroxyalkylamine, carboxylalkylamine,        alkylcarboxylalkylamine, arylamine, alkyl, alkenyl, aryl,        substituted alkyl, substituted alkenyl, or substituted aryl;    -   Y is hydrogen, hydroxy, alkoxy, or protected hydroxy; and    -   the dashed line represents an optional double bond.

Other aspects and features of the invention are detailed below.

DESCRIPTION OF THE FIGURES

FIG. 1A presents TLR2 antagonist screening in which TLR2 was stimulatedwith HKLM at 10⁸ cell/ml. Plotted is the activity of a secreted alkalinephosphatase reporter system in optical density (OD) at 650 nm for eachtreatment condition, which was tested in duplicate. All agents weretested at 10 μM.

FIG. 1B presents TLR3 antagonist screening in which TLR3 was stimulatedwith poly(I:C) at 1 μg/ml. Plotted is the activity of a secretedalkaline phosphatase reporter system in optical density (OD) at 650 nmfor each treatment condition, which was tested in duplicate. All agentswere tested at 10 μM.

FIG. 1C presents TLR4 antagonist screening in which TLR4 was stimulatedwith LPS at 100 ng/ml. Plotted is the activity of a secreted alkalinephosphatase reporter system in optical density (OD) at 650 nm for eachtreatment condition, which was tested in duplicate. All agents weretested at 10 μM.

FIG. 1D presents TLR5 antagonist screening in which TLR5 was stimulatedwith Flagellin at 100 ng/ml. Plotted is the activity of a secretedalkaline phosphatase reporter system in optical density (OD) at 650 nmfor each treatment condition, which was tested in duplicate. All agentswere tested at 10 μM.

FIG. 1E presents TLR7 antagonist screening in which TLR7 was stimulatedwith CL097 at 1 μg/ml. Plotted is the activity of a secreted alkalinephosphatase reporter system in optical density (OD) at 650 nm for eachtreatment condition, which was tested in duplicate. All agents weretested at 10 μM.

FIG. 1F presents TLR8 antagonist screening in which TLR8 was stimulatedwith CL075 at 1 μg/ml. Plotted is the activity of a secreted alkalinephosphatase reporter system in optical density (OD) at 650 nm for eachtreatment condition, which was tested in duplicate. All agents weretested at 10 μM.

FIG. 1G presents TLR9 antagonist screening in which TLR9 was stimulatedwith CpG ODN 2006 at 100 ng/ml. Plotted is the activity of a secretedalkaline phosphatase reporter system in optical density (OD) at 650 nmfor each treatment condition, which was tested in duplicate. All agentswere tested at 10 μM.

FIG. 2A illustrates the analgesic effects of (+)-naloxone on mechanicalallodynia in rats. Plotted is 50% allodynia threshold at threetimepoints for each treatment group on Day 14 for the left (affected)paw. Bars represent mean+/−SEM. *p<0.05, **p<0.01, ***p<0.001 vs.vehicle.

FIG. 2B presents the analgesic effects of (+)-naloxone on mechanicalallodynia in rats. Plotted is 50% allodynia threshold at three timepoints for each treatment group on Day 14 for the right (unaffected)paw. Bars represent mean+/−SEM. *p<0.05, **p<0.01, ***p<0.001 vs.vehicle.

FIG. 3A depicts development of allodynia. Plotted is the von Frey scoreon days 1, 4 and 14 for ipsilateral paws in each treatment group.

FIG. 3B depicts development of allodynia. Plotted is the von Frey scoreon days 1, 4 and 14 for contralateral paws in each treatment group.

FIG. 4 presents allodynia responses prior to and 3 hrs after treatmentwith various doses of compound II-25. Panels A-D present responses inipsilateral paws. Panels E-H present responses in contralateral paws.

FIG. 5A presents area under the curve (AUC) data for each group treatedwith compound II-25 in ipsilateral paws.

FIG. 5B presents area under the curve (AUC) data for each group treatedwith compound II-25 in contralateral paws.

FIG. 6 depicts allodynia responses prior to and 3 hrs after treatmentwith various doses of compound II-93. Panels A-D present responses inipsilateral paws. Panels E-H present responses in contralateral paws.

FIG. 7A presents area under the curve (AUC) data for each group treatedwith compound II-93 in ipsilateral paws.

FIG. 7B presents area under the curve (AUC) data for each group treatedwith compound II-93 in contralateral paws.

FIG. 8 illustrates that (+)-naloxone did not affectacetaminophen-induced hepatotoxicity. Panel A presents liver histologyscores and panel B presents alanine transaminase (ALT) activity incontrol and treated groups.

FIG. 9 illustrates that compound II-78 provided protection foracetaminophen-induced hepatotoxicity. Panel A presents liver histologyscores and panel B presents alanine transaminase (ALT) activity incontrol and treated groups.

FIG. 10 presents a time course of clinical scores of experimentalautoimmune encephalomyelitis following treatment with a variety of(+)-morphinans.

FIG. 11 shows that compound II-78 (COV) reduced hepatic ischemicreperfusion (IR) injury. Plotted is the average histological Suzukiscore under the indicated conditions after 3 hours of perfusion (panelA) and 12 hours of perfusion (panel B). Statistically significantdifferences are indicated as: * p<0.05, and ** p<0.001.

FIG. 12 illustrates that compound II-78 (COV) reduced IR-induced serumalanine transaminase (ALT) levels. Plotted is serum ALT activity (U/L)after 3 hours of perfusion (panel A) and 12 hours of perfusion (panelB). Panel C presents a time course of serum ALT activity under theindicated conditions. Statistically significant differences areindicated as: * p<0.05, and ** p<0.001.

FIG. 13 shows that compound II-78 (COV) reduced IR-inducedpro-inflammatory cytokines and increased an anti-inflammatory moleculeafter 12 hours of perfusion. Plotted is the level of mRNA relative toβ-actin for IL-1β (panel A), MCP-1 (panel B), NLPR3 (panel C), andIL-1Ra (panel D). Statistically significant differences are indicatedas: * p<0.05, ** p<0.01, and *** p<0.001.

FIG. 14 documents the reduction of IR-induced activation of MAP kinasesby compound II-78 (COV) (panel A) and the reduction of IR-inducedcleavage and activation of LTR9 compound II-78 (COV) (panel B), asassessed by western blotting.

FIG. 15 shows that compound II-78 (COV) compound reduced both NFκB- andIRF-dependent pathways downstream of TLR9. Plotted is the level of mRNArelative to β-actin for IL-1β (panel A) and IFN-β (panel B).Statistically significant differences are indicated as: * p<0.05, **p<0.01, and *** p<0.001.

FIG. 16 documents that compound II-78 (COV) reduced hepatic ischemicreperfusion (IR) injury in steatotic livers. Plotted is the averagehistological Suzuki score under the indicated conditions after 3 hoursof perfusion (panel A) and 12 hours of perfusion (panel B).Statistically significant differences are indicated as: * p<0.05, and **p<0.001.

FIG. 17 illustrates that compound II-78 (COV) reduced IR-induced serumalanine transaminase (ALT) levels in steatotic livers. Plotted is serumALT activity (U/L) after 3 hours of perfusion (panel A) and 12 hours ofperfusion (panel B). Panel C presents a time course of serum ALTactivity under the indicated conditions. Statistically significantdifferences are indicated as: * p<0.05, and ** p<0.001.

FIG. 18 shows that compound II-78 (COV) reduced IR-inducedpro-inflammatory cytokines and increased an anti-inflammatory moleculeafter 12 hours of perfusion. Plotted is the level of mRNA relative toβ-actin for IL-1β (panel A), MCP-1 (panel B), NLPR3 (panel C), andIL-1Ra (panel D). Statistically significant differences are indicatedas: * p<0.05, ** p<0.01, and *** p<0.001.

FIG. 19 documents the reduction of IR-induced activation of MAP kinasesby compound II-78 (COV) (panel A) and the reduction of IR-inducedcleavage and activation of LTR9 by compound II-78 (COV) (panel B), asassessed by western blotting.

FIG. 20 shows the effects of compound II-78 (COV08-0064) on a model ofacute pancreatitis. The compound reduced neutrophil infiltration in thepancreas (panel A), pancreatic trypsin activity (panel B), reduced serumamylase (panel C), and reduced pancreatic caspase 1 activity (panel D).

DETAILED DESCRIPTION OF THE INVENTION

It has been discovered that certain (+)-morphinans block the activationof TLR9 and, consequently, the activation of glial cells. Thus,(+)-morphinans comprising TLR9 antagonist activity may be used to treatpain, as well as other conditions associated with pain and inflammation.It has also been discovered that the inhibition of TLR9 activation maybe used as a screening tool to identify (+)-morphinans that may betherapeutically effective in treating conditions such as traumatic pain,neuropathic pain, inflammatory disorders, acetaminophen toxicity,autoimmune disorders, neurodegenerative disorders, and cancer.Accordingly, the present invention provides (+)-morphinans comprisingTLR9 antagonist activity, methods for inhibiting the activation of TLR9,screening methods for identifying therapeutically effective(+)-morphinans, and methods of using the (+)-morphinans comprising TLR9antagonist activity to treat conditions such as traumatic pain,neuropathic pain, inflammatory disorders, acetaminophen toxicity,autoimmune disorders, neurodegenerative disorders, and cancer.

(I) (+)-Morphinans Comprising TLR9 Antagonist Activity

(a) Compounds Comprising Formula (I)

One aspect of the present invention is the provision of (+)-morphinanscomprising TLR9 antagonist activity. In one embodiment, the(+)-morphinan may be a compound comprising Formula (I) or apharmaceutically acceptable salt thereof:

wherein:

-   -   A is chosen from {−}C(═O){−}, {−}C(S){−}, {−}C(═CH₂){−},        {−}CH(A₁){−}, and {−}C(A₁)(A₂){−};    -   A₁ and A₂ are independently chosen from hydrogen, alkyl,        alkenyl, alkoxy, acyloxy, aryl, heteroaryl, hydroxy,        hydroxyalkyl, polyhydroxyalkyl, amine, and amide, wherein when        both A₁ and A₂ are present, together they may form a carbocyclic        ring or heterocyclic ring;    -   R and R′ are independently chosen from hydrogen, hydroxy, amine,        hydrocarbyl, and substituted hydrocarbyl, wherein R′ is        optional, as represented by the dashed line;    -   R¹, R², R³, R⁴, R⁵, R⁶, R⁷, R⁸, R⁹, R¹⁰, R¹¹, and R¹² are        independently chosen from hydrogen, hydroxy, amine, halo,        hydrocarbyl, and substituted hydrocarbyl, wherein R⁷ and A₁ may        together form a ring or a ring system chosen from carbocyclic,        heterocyclic, aryl, heteroaryl, and combinations thereof;    -   Y is chosen from hydrogen, hydroxy, alkoxy, acyloxy, amine, and        amide; and    -   the dashed lines between the carbons atoms at positions 5 and 6,        6 and 7, 7 and 8, and 8 and 14 represent carbon-carbon single        bonds, carbon-carbon double bonds, or combinations thereof,        provided that if there is a double bond between the carbons at        positions 5 and 6 then only one of R⁵ or R⁶ is present, if there        is a double bond between the carbons at 6 and 7 then only one of        R⁷ or R⁸ is present, if there is a double bond between the        carbons at 7 and 8 then only one of R⁷ or R⁸ is present and only        one of R⁹ or R¹⁰ is present, and if there is a double bond        between the carbons at 8 and 14 then only one of R⁹ or R¹⁰ is        present and Y is not present.

In one iteration of this embodiment, R, R¹, R², R³, R⁴, R⁵, R⁶, R⁷, R⁸,R⁹, R¹⁰, R¹¹, and R¹² are independently chosen from hydrogen, hydroxy,alkyl, alkenyl, alkynyl, aminoalkyl, alkoxyalkyl, aralkyl, cycloalkyl,hydroxyalkyl, acyloxy, alkoxy, haloalkoxyl, aryl, amine, amide, andhalo.

In another iteration, R², R⁶, R⁸, R⁹, R¹⁰, R¹¹, and R¹² are hydrogen; Ris chosen from hydrogen, methyl, alkyl, alkenyl, allyl,methylcycloalkyl, methylcyclopropyl, methylcyclobutyl, methylaryl,methylphenyl, acyl, acylalkyl, acylcycloalkyl, acylcyclopropyl,acylcyclobutyl, acylaryl, acylphenyl, acyloxy, acyloxyalkyl,acyloxyaryl, acyloxyphenyl, alkoxy, and alkoxyalkyl; R¹ is chosen fromhydrogen, halo, alkyl, alkenyl, alkoxyalkyl, alkoxyalkenyl, aryl,heteroaryl, and furanyl; R³ and R⁴ are independently chosen fromhydroxy, alkoxy, methoxy, acyloxy, and protected hydroxy; R⁷ is chosenfrom hydroxy, alkoxy, methoxy, acyloxy, protected hydroxy, hydrocarbyl,and substituted hydrocarbyl, wherein R⁷ and A¹ may together form anindolyl ring; and Y is hydrogen or hydroxy.

(b) Compounds Comprising Formula (II)

In another embodiment, the (+)-morphinan may be a compound comprisingFormula (II) or a pharmaceutically acceptable salt thereof:

wherein:

-   -   A is chosen from {−}C(═O){−}, {−}C(S){−}, {−}C(═CH₂){−},        {−}CH(A₁){−}, and {−}C(A₁)(A₂){−};    -   A₁ and A₂ are independently chosen from hydrogen, alkyl,        alkenyl, alkoxy, acyloxy, aryl, heteroaryl, hydroxy,        hydroxyalkyl, polyhydroxyalkyl, amine, and amide, wherein when        both A₁ and A₂ are present, together they may form a carbocyclic        ring or heterocyclic ring;    -   R and R′ are independently chosen from hydrogen, hydroxy, amine,        hydrocarbyl, and substituted hydrocarbyl, wherein R′ is        optional, as represented by the dashed line;    -   R¹, R², R³, R⁷, R⁸, R⁹, R¹⁰, R¹¹, and R¹² are independently        chosen from hydrogen, hydroxy, amine, halo, hydrocarbyl, and        substituted hydrocarbyl, wherein R⁷ and A¹ may together form a        ring or a ring system chosen from carbocyclic, heterocyclic,        aryl, heteroaryl, and combinations thereof;    -   Y is chosen from hydrogen, hydroxy, alkoxy, acyloxy, amine, and        amide;    -   the dashed lines between the carbons atoms at positions 6 and 7,        7 and 8, and 8 and 14 represent carbon-carbon single bonds,        carbon-carbon double bonds, or combinations thereof, provided        that if there is a double bond between the carbons at 6 and 7        then only one of R⁷ or R⁸ is present, if there is a double bond        between the carbons at 7 and 8 then only one of R⁷ or R⁸ is        present and only one of R⁹ or R¹⁰ is present, and if there is a        double bond between the carbons at 8 and 14 then only one of R⁹        or R¹⁰ is present and Y is not present; and    -   the carbons at positions 6 and 14 may be linked by a moiety        chosen from ether, alkyl, alkenyl, substituted alkyl, and        substituted alkenyl.

In one iteration of this embodiment, R, R¹, R², R³, R⁷, R⁸, R⁹, R¹⁰,R¹¹, and R¹² are independently chosen from hydrogen, hydroxy, alkyl,alkenyl, alkynyl, aminoalkyl, alkoxyalkyl, aralkyl, cycloalkyl,hydroxyalkyl, acyloxy, alkoxy, haloalkoxyl, aryl, amine, amide, andhalo.

In another iteration, R², R⁸, R⁹, R¹⁰, R¹¹, and R¹² are hydrogen; R ischosen from hydrogen, methyl, alkyl, alkenyl, allyl, methylcycloalkyl,methylcyclopropyl, methylcyclobutyl, methylaryl, methylphenyl, acyl,acylalkyl, acylcycloalkyl, acylcyclopropyl, acylcyclobutyl, acyl aryl,acylphenyl, acyloxy, acyloxyalkyl, acyloxyaryl, acyloxyphenyl, alkoxy,and alkoxyalkyl; R¹ is chosen from hydrogen, halo, alkyl, alkenyl,alkoxyalkyl, alkoxyalkenyl, aryl, heteroaryl, and furanyl; R³ is chosenfrom hydroxy, alkoxy, methoxy, acyloxy, and protected hydroxy; R⁷ ischosen from hydroxy, alkoxy, methoxy, acyloxy, protected hydroxy,hydrocarbyl, and substituted hydrocarbyl, wherein R⁷ and A₁ may togetherform an indolyl ring; and Y is hydrogen or hydroxy.

In preferred embodiments, the (+)-morphinan may be a compound comprisingFormula (IIa) or a pharmaceutically acceptable salt thereof:

wherein:

-   -   R is chosen from hydrogen, alkyl, alkenyl, and methylcycloalkyl;    -   R¹ is chosen from hydrogen, aryl, and heteroaryl;    -   R³ is chosen from hydroxy, alkoxy, and aryloxy;    -   A₁ is chosen from hydroxy, keto, C₂-C₆ alkoxy, acyloxy, aryloxy,        amino, alkylamine, hydroxyalkylamine, carboxylalkylamine,        alkylcarboxylalkylamine, and arylamine; provided, however, that        when A₁ is keto, R¹ is other than hydrogen; when A₁ is hydroxy,        R is other than methyl or allyl; when A₁ is hydroxy, Y is other        than hydroxy; or when A₁ is an amine, Y is other than hydroxy;    -   Y is chosen from hydrogen and hydroxy; and    -   the dashed line represents an optional double bond.

In one exemplary embodiment, the (+)-morphinan may be a compoundcomprising Formula (IIb) or a pharmaceutically acceptable salt thereof:

wherein:

-   -   R is chosen from hydrogen, methyl, allyl, methylcyclopropyl, and        methylcyclobutyl;    -   R^(a) is chosen from hydrogen, methyl, ethyl, phenyl, and        benzyl;    -   R^(b) is chosen from hydrogen, methyl, alkyl, alkylalcohol, and        alkylcarboxylalkylester; and    -   Y is hydrogen.

In one iteration, R is methyl, R^(a) is methyl, R^(b) ismethylacetylmethylester; and Y is hydrogen. In this iteration,therefore, the (+)-morphinan is compound II-92 in Table 2. In anotheriteration, R is allyl, R^(a) is methyl, R^(b) is propyl; and Y ishydrogen. That is, the (+)-morphinan is compound II-93 in Table 2. In afurther iteration, R is allyl, R^(a) is methyl, R^(b) is ethylalcohol;and Y is hydrogen. Thus, the (+)-morphinan is compound II-21 in Table 2.

In another exemplary embodiment, the (+)-morphinan may be a compoundcomprising Formula (IIc) or a pharmaceutically acceptable salt thereof:

wherein:

-   -   R is chosen from hydrogen, methyl, allyl, methylcyclopropyl, and        methylcyclobutyl;    -   R¹ is chosen from hydrogen, aryl, heteroaryl, and furanyl;    -   R^(a) is chosen from hydrogen, methyl, ethyl, phenyl, and        benzyl;    -   R^(c) is chosen from hydroxy, C₂-C₆ alkoxy, and keto; provided,        however, that when R^(c) is keto, R¹ is other than hydrogen;        when R^(c) is hydroxy, R is other than methyl or allyl; or when        R^(c) is hydroxy, Y is other than hydroxy;    -   Y is chosen from hydrogen and hydroxy; and    -   the dashed line represents an optional double bond.

In one iteration of this embodiment, R is methylcyclopropyl, R¹ ishydrogen, R^(a) is methyl, R^(c) is hydroxy; and Y is hydrogen. That is,the (+)-morphinan is compound II-25 in Table 2. In yet anotheriteration, R is allyl, R¹ is furanyl, R^(a) is methyl, R^(c) is keto;and Y is hydrogen. Thus, the (+)-morphinan is compound II-2 in Table 2.

(c) Compounds Comprising Formula (III)

In still another embodiment, the (+)-morphinan having TLR9 antagonistactivity may be a compound comprising Formula (III) or apharmaceutically acceptable salt thereof:

wherein:

-   -   R and R′ are independently chosen from hydrogen, hydroxy, amine,        hydrocarbyl, and substituted hydrocarbyl, wherein R′ is        optional, as represented by the dashed line; and    -   R¹, R², R³, R⁴, R⁵, R⁶, R⁷, R⁸, R⁹, R¹⁰, R¹¹, and R¹² are        independently chosen from hydrogen, hydroxy, amine, halo,        hydrocarbyl, and substituted hydrocarbyl.

In one iteration of this embodiment, R, R¹, R², R³, R⁴, R⁵, R⁶, R⁷, R⁸,R⁹, R¹⁰, R¹¹, and R¹² are independently chosen from hydrogen, hydroxy,alkyl, alkenyl, alkynyl, aminoalkyl, alkoxyalkyl, aralkyl, cycloalkyl,hydroxyalkyl, acyloxy, alkoxy, haloalkoxyl, aryl, amine, amide, andhalo.

In another iteration, R¹, R², R⁵, R⁶, R⁸, R⁹, R¹⁰, R¹¹, and R¹² arehydrogen; R is chosen from hydrogen, methyl, alkyl, alkenyl, allyl,methylcycloalkyl, methylcyclopropyl, methylcyclobutyl, methylaryl,methylphenyl, acyl, acylalkyl, acylcycloalkyl, acylcyclopropyl,acylcyclobutyl, acylaryl, acylphenyl, acyloxy, acyloxyalkyl,acyloxyaryl, acyloxyphenyl, alkoxy, and alkoxyalkyl; and R³, R⁴, and R⁷are independently chosen from hydroxy, alkoxy, methoxy, acyloxy, andprotected hydroxy.

In an exemplary embodiment, the (+)-morphinan may be a compoundcomprising Formula (IIIa) or a pharmaceutically acceptable salt thereof:

wherein:

-   -   R is chosen from C₂-C₆ alkyl, alkenyl, methylcycloalkyl,        methylaryl, acylalkyl, acylcycloalkyl, acylaryl, acyloxy,        acyloxyalkyl, acyloxyaryl, alkoxy, and alkoxyalkyl; and    -   R³, R⁴, and R⁷ are independently chosen from hydroxy, alkoxy,        aryloxy, and acyloxy.

In one iteration of this embodiment, R is allyl, R³ is methoxy, R⁴ ishydroxy, and R⁷ is methoxy. That is, the (+)-morphinan is compound III-2in Table 2.

(d) Pharmaceutically Acceptable Salts

Any of the compounds detailed above in sections (I)(a), (I)(b), and(I)(c) may be provided as a pharmaceutically acceptable salt. The term“pharmaceutically acceptable salt” refers to a salt commonly used toform an alkali metal salt or addition salt of a free acid or a freebase. The nature of the salt may vary, provided that it ispharmaceutically acceptable. Suitable pharmaceutically acceptable acidaddition salts of compounds of the present invention may be preparedfrom an inorganic acid or from an organic acid. Examples of suchinorganic acids are hydrochloric, hydrobromic, hydroiodic, nitric,carbonic, sulfuric and phosphoric acid. Appropriate organic acids may beselected from aliphatic, cycloaliphatic, aromatic, araliphatic,heterocyclic, carboxylic and sulfonic classes of organic acids, examplesof which are formic, acetic, propionic, succinic, glycolic, gluconic,lactic, malic, tartaric, citric, ascorbic, glucuronic, maleic, fumaric,pyruvic, aspartic, glutamic, benzoic, anthranilic, mesylic,4-hydroxybenzoic, phenylacetic, mandelic, embonic (pamoic),methanesulfonic, ethanesulfonic, benzenesulfonic, pantothenic,2-hydroxyethanesulfonic, toluenesulfonic, sulfanilic,cyclohexylaminosulfonic, stearic, algenic, hydroxybutyric, salicylic,galactaric and galacturonic acid. Suitable pharmaceutically-acceptablebase addition salts of compounds of the present invention includemetallic salts made from aluminum, calcium, lithium, magnesium,potassium, sodium and zinc or organic salts made from N,N′-dibenzylethylenediamine, chloroprocaine, choline, diethanolamine,ethylenediamine, meglumine (i.e., N-methylglucamine), and procaine. Allof these salts may be prepared by conventional means from thecorresponding compound by reacting, for example, the appropriate acid orbase with the any of the compounds of the invention.

(e) Stereochemistry

Each of the compounds detailed above in sections (I)(a), (I)(b), and(I)(c) has a (+) orientation with respect to the rotation of polarizedlight. More specifically, each chiral carbon has an R or an Sconfiguration. As will be appreciated by a skilled artisan, the R or Sconfiguration for a given carbon may change depending on the particularstructure of the compound and the compound's substitution pattern.

(II) Pharmaceutical Compositions Comprising a (+)-Morphinan and aCompound Comprising Formula (IV)

Still another aspect of the present invention provides a pharmaceuticalcomposition comprising a (+)-morphinan or a pharmaceutically acceptablesalt thereof and a compound comprising Formula (IV) or apharmaceutically acceptable salt thereof, provided that the(+)-morphinan is other than dextrorphan or a derivative thereof.

(a) Compounds Comprising Formula (IV)

The pharmaceutical composition of the invention comprises a compoundcomprising Formula (IV) or a pharmaceutically acceptable salt thereof.In general, the compound comprising Formula (IV) may have weak μ-opioidreceptor agonist activity and/or monoamine reuptake inhibitor activity.The compound comprising Formula (IV) has the following structure:

wherein:

-   -   R²¹ is chosen from hydrogen, hydrocarbyl, and substituted        hydrocarbyl;    -   R²² is chosen from hydrocarbylamine and substituted        hydrocarbylamine; and    -   Z is chosen from oxygen and sulfur.

In preferred embodiments, R²¹ is chosen from hydrogen, alkyl, andsubstituted alkyl; R²² is chosen from dialkylaminoalkyl and substituteddialkylaminoalkyl; and Z is oxygen. In one preferred embodiment, R²¹ ischosen from hydrogen and methyl; and R²² is chosen fromdimethylamino-ethyl-methylpropyl and dimethylamino cyclohexanol. In onepreferred embodiment, R²¹ is hydrogen, R²² isdimethylamino-ethyl-methylpropyl, and Z is oxygen (i.e., the compound istapentadol). In another preferred embodiment, R²¹ is methyl, R²² isdimethylamino cyclohexanol, and Z is oxygen (i.e., the compound istramadol).

The compound comprising Formula (IV) may be provided as apharmaceutically acceptable salt. The pharmaceutically acceptable saltmay be an acid addition salt or a base addition salt. Acids commonlyemployed to form acid addition salts include inorganic acids, such asfor example, hydrochloric acid, hydrobromic acid, hydroiodic acid,sulfuric acid, phosphoric acid, and the like, and organic acids such asp-toluenesulfonic, methanesulfonic acid, oxalic acid,p-bromophenylsulfonic acid, carbonic acid, succinic acid, citric acid,benzoic acid, acetic acid, and the like. Examples of suchpharmaceutically acceptable salts are the sulfate, pyrosulfate,bisulfate, sulfite, bisulfite, phosphate, monohydrogenphosphate,dihydrogenphosphate, metaphosphate, pyrophosphate, chloride, bromide,iodide, acetate, propionate, decanoate, caprylate, acrylate, formate,isobutylate, caproate, heptanoate, propiolate, oxalate, malonate,succinate, suberate, sebacate, fumarate, maleate, butyne-1,4-dioate,hexyne-1,6-dioate, benzoate, chlorobenzoate, methylbenzoate,dinitrobenzoate, hydroxybenzoate, methoxybenzoate, phthalate, sulfonate,xylenesulfonate, phenylacetate, phenylpropionate, phenylbutylate,citrate, lactate, g-hydroxybutylate, glycolate, tartrate,methanesulfonate, propanesulfonate, naphthalene-1-sulfonate,napththalene-2-sulfonate, mandelate and the like. Base addition saltsinclude those derived from inorganic bases, such as for example,ammonium or alkali or alkaline earth metal hydroxides, carbonates,bicarbonates, and the like. Such bases include sodium hydroxide,potassium hydroxide, ammonium hydroxide, potassium carbonate, sodiumcarbonate, sodium bicarbonate, potassium bicarbonate, calcium hydroxide,and calcium carbonate.

The compound comprising Formula (IV) may be a (+) enantiomer, a (−)enantiomer, or a combination thereof. Thus, each chiral carbon may havean R or an S configuration.

(b) (+)-Morphinan Compound

The pharmaceutical composition of the invention also comprises a(+)-morphinan or a pharmaceutically acceptable salt thereof.

(i) Compounds Comprising Formula (I)

In one embodiment, the (+)-morphinan may be a compound comprisingFormula (I) or a pharmaceutically acceptable salt thereof, as detailedabove in section (I)(a); provided however that when A₁ is present, it isother than hydrogen, or when both A₁ and A₂ are present, each is otherthan hydrogen. Stated another way, the compound comprising Formula (I)is other than dextrorphan or a derivative thereof.

(ii) Compounds Comprising Formula (II)

In another embodiment, the (+)-morphinan may be a compound comprisingFormula (II) or a pharmaceutically acceptable salt thereof, as detailedabove in section (I)(b).

In preferred embodiments, the (+)-morphinan may be a compound comprisingFormula (IIa) or a pharmaceutically acceptable salt thereof:

wherein;

-   -   R is chosen from hydrogen, methyl, alkyl, alkenyl, allyl,        methylcycloalkyl, methylcyclopropyl, methylcyclobutyl,        methylaryl, methylphenyl, acyl, acylalkyl, acylcycloalkyl,        acylcyclopropyl, acylcyclobutyl, acylaryl, acylphenyl, acyloxy,        acyloxyalkyl, acyloxyaryl, acyloxyphenyl, alkoxy, and        alkoxyalkyl;    -   R¹ is chosen from hydrogen, halo, hydroxy, alkoxy, alkoxyalkyl,        alkoxyalkenyl, aryloxy, alkyl, alkenyl, aryl, heteroaryl, and        amine;    -   R³ is chosen from hydroxy, alkoxy, aryloxy, acyl, acyloxy, and        protected hydroxy;    -   A₁ is chosen from hydrogen, hydroxy, keto, alkoxy, acyl,        acyloxy, amino, amide, alkylamine, hydroxyalkylamine,        carboxylalkylamine, alkylcarboxylalkylamine, arylamine, alkyl,        alkenyl, aryl, substituted alkyl, substituted alkenyl, and        substituted aryl;    -   Y is chosen from hydrogen, hydroxy, alkoxy, and protected        hydroxy; and    -   the dashed line represents an optional double bond.

In one exemplary embodiment, the (+)-morphinan may be a compoundcomprising Formula (IIb) or a pharmaceutically acceptable salt thereof:

wherein:

-   -   R is chosen from hydrogen, alkyl, methyl, allyl,        methylcyclopropyl, and methylcyclobutyl;    -   R^(a) is chosen from hydrogen, methyl, alkyl, phenyl, and        benzyl;    -   R^(b) is chosen from hydrogen, methyl, alkyl, alkylalcohol,        alkylcarboxyl, alkylcarboxylalkylester, acyl, acylalkyl,        acylaryl, acyloxy, acyloxyalkyl, acyloxyaryl; and    -   Y is chosen from hydrogen and hydroxy.

In one iteration of this embodiment, R is methylcyclopropyl, R^(a) ishydrogen, R^(b) is methyl, and Y is hydroxy. In another iteration, R ismethyl, R^(a) is methyl, R^(b) is methylacetylmethylester; and Y ishydrogen. In yet another iteration, R is allyl, R^(a) is methyl, R^(b)is propyl; and Y is hydrogen. In a further iteration, R is allyl, R^(a)is methyl, R^(b) is ethylalcohol; and Y is hydrogen.

In another exemplary embodiment, the (+)-morphinan may be a compoundcomprising Formula (IIc) or a pharmaceutically acceptable salt thereof:

wherein:

-   -   R is chosen from hydrogen, methyl, alkyl, allyl,        methylcyclopropyl, and methylcyclobutyl;    -   R¹ is chosen from hydrogen, halo, hydroxy, alkoxy, amine, aryl,        heteroaryl, and furanyl;    -   R^(a) is chosen from hydrogen, methyl, alkyl, phenyl, and        benzyl;    -   R^(c) is chosen from hydroxy, methoxy, alkoxy, keto, and acyl;    -   Y is chosen from hydrogen and hydroxy; and    -   the dashed line represents an optional double bond.

In one iteration of this embodiment, R is methylcyclopropyl, R¹ ishydrogen, R^(a) is methyl, R^(c) is hydroxy; and Y is hydrogen. Inanother iteration, R is methylcyclopropyl, R¹ is hydrogen, R^(a) ismethoxy, R^(c) is methyl, Y is hydrogen, and the optional double bond ispresent. In a further iteration, R is allyl, R¹ is furanyl, R^(a) ismethyl, R^(c) is keto, and Y is hydrogen. In still another iteration, Ris hydrogen, R¹ is hydrogen, R^(a) is methyl, R^(c) is methoxy, Y ishydrogen, and the optional double bond is present.

(iii) Compounds Comprising Formula (III)

In another embodiment, the (+)-morphinan may be a compound comprisingFormula (III) or a pharmaceutically acceptable salt thereof, as detailedabove in section (I)(c).

In an exemplary embodiment, the (+)-morphinan may be a compoundcomprising Formula (IIIa) or a pharmaceutically acceptable salt thereof:

wherein:

-   -   R is chosen from hydrogen, methyl, alkyl, alkenyl, allyl,        methylcycloalkyl, methylcyclopropyl, methylcyclobutyl,        methylaryl, methylphenyl, acyl, acylalkyl, acylcycloalkyl,        acylcyclopropyl, acylcyclobutyl, acylaryl, acylphenyl, acyloxy,        acyloxyalkyl, acyloxyaryl, acyloxyphenyl, alkoxy, and        alkoxyalkyl; and    -   R³, R⁴, and R⁷ are independently chosen from hydrogen, hydroxy,        alkoxy, aryloxy, acyloxy, amine, halo, and protected hydroxy

In one iteration of this embodiment, R is allyl, R³ is methoxy, R⁴ ishydroxy, and R⁷ is methoxy.

(c) Optional Excipients

The pharmaceutical composition may further comprise at least onepharmaceutically acceptable excipient. Non-limiting examples of suitableexcipients include diluents, binders, fillers, buffering agents, pHmodifying agents, effervescent disintegrants, non-effervescentdisintegrants, dispersing agents, stabilizers, preservatives, compactionagents, lubricants, coloring agents and/or flavoring agents. The amountand types of excipients utilized to form the pharmaceutical compositionmay be selected according to known principles of pharmaceutical science.

In one embodiment, the excipient may include at least one diluent. Thediluent may be compressible (i.e., plastically deformable) or abrasivelybrittle. Non-limiting examples of suitable compressible diluents includemicrocrystalline cellulose (MCC), cellulose derivatives, cellulosepowder, cellulose esters (i.e., acetate and butyrate mixed esters),ethyl cellulose, methyl cellulose, hydroxypropyl cellulose,hydroxypropyl methylcellulose, sodium carboxymethylcellulose, cornstarch, phosphated corn starch, pregelatinized corn starch, rice starch,potato starch, tapioca starch, starch-lactose, starch-calcium carbonate,sodium starch glycolate, glucose, fructose, lactose, lactosemonohydrate, sucrose, xylose, lacitol, mannitol, malitol, sorbitol,xylitol, maltodextrin, and trehalose. Non-limiting examples of suitableabrasively brittle diluents include dibasic calcium phosphate (anhydrousor dihydrate), calcium phosphate tribasic, calcium carbonate, andmagnesium carbonate.

In another embodiment, the excipient may comprise a binder. Suitablebinders include, but are not limited to, starches, pregelatinizedstarches, gelatin, polyvinylpyrrolidone, cellulose, methylcellulose,sodium carboxymethylcellulose, ethylcellulose, polyacrylamides,polyvinyloxoazolidone, polyvinylalcohols, C₁₂-C₁₈ fatty acid alcohol,polyethylene glycol, polyols, saccharides, oligosaccharides,polypeptides, oligopeptides, and combinations thereof.

In another embodiment, the excipient may include a filler. Suitablefillers include, but are not limited to, carbohydrates, inorganiccompounds, and polyvinylpyrrolidone. By way of non-limiting example, thefiller may be calcium sulfate, both di- and tri-basic, starch, calciumcarbonate, magnesium carbonate, microcrystalline cellulose, dibasiccalcium phosphate, magnesium carbonate, magnesium oxide, calciumsilicate, talc, modified starches, lactose, sucrose, mannitol, orsorbitol.

In still another embodiment, the excipient may comprise a bufferingagent. Representative examples of suitable buffering agents include, butare not limited to, MOPS, HEPES, TAPS, Bicine, Tricine, TES, PIPES, MES,Tris buffers or buffered saline salts (e.g., Tris buffered saline orphosphate buffered saline).

In various embodiments, the excipient may include a pH modifier. By wayof non-limiting example, the pH modifying agent may be citric acid,sodium carbonate, or sodium bicarbonate.

In a further embodiment, the excipient may include a non-effervescentdisintegrant. Suitable examples of non-effervescent disintegrantsinclude, but are not limited to, starches such as corn starch, potatostarch, pregelatinized and modified starches thereof, sweeteners, clays,such as bentonite, micro-crystalline cellulose, alginates, sodium starchglycolate, gums such as agar, guar, locust bean, karaya, pecitin, andtragacanth.

In another embodiment, the excipient may comprise an effervescentdisintegrant. By way of non-limiting example, suitable effervescentdisintegrants include sodium bicarbonate in combination with citric acidand sodium bicarbonate in combination with tartaric acid.

In another alternate embodiment, the excipient may also include apreservative. Non-limiting examples of suitable preservatives includeantioxidants, such as alpha-tocopherol or ascorbate, and antimicrobials,such as parabens, chlorobutanol or phenol.

In yet another embodiment, the excipient may include a dispersionenhancer. Suitable dispersants may include, but are not limited to,starch, alginic acid, polyvinylpyrrolidones, guar gum, kaolin,bentonite, purified wood cellulose, sodium starch glycolate,isoamorphous silicate, and microcrystalline cellulose as high HLBemulsifier surfactants.

In a further embodiment, the excipient may include a lubricant.Non-limiting examples of suitable lubricants include minerals such astalc or silica; and fats such as vegetable stearin, magnesium stearateor stearic acid.

In still another embodiment, it may be desirable to provide a coloringagent. Suitable color additives include, but are not limited to, food,drug and cosmetic colors (FD&C), drug and cosmetic colors (D&C), orexternal drug and cosmetic colors (Ext. D&C).

In a further embodiment, the excipient may include flavoring agents.Flavoring agents may be chosen from synthetic flavor oils and flavoringaromatics and/or natural oils, extracts from plants, leaves, flowers,fruits, and combinations thereof. By way of example, these may includecinnamon oils, oil of wintergreen, peppermint oils, clover oil, hay oil,anise oil, eucalyptus, vanilla, citrus oils (such as lemon oil, orangeoil, grape and grapefruit oil), and fruit essences (such as apple,peach, pear, strawberry, raspberry, cherry, plum, pineapple, andapricot). In still another embodiment, the excipient may include asweetener. By way of non-limiting example, the sweetener may be selectedfrom glucose (corn syrup), dextrose, invert sugar, fructose, andmixtures thereof (when not used as a carrier); saccharin and its varioussalts such as the sodium salt; dipeptide sweeteners such as aspartame;dihydrochalcone compounds, glycyrrhizin; stevia-derived sweeteners;chloro derivatives of sucrose such as sucralose; sugar alcohols such assorbitol, mannitol, sylitol, and the like. Also contemplated arehydrogenated starch hydrolysates and the synthetic sweetener3,6-dihydro-6-methyl-1,2,3-oxathiazin-4-one-2,2-dioxide, particularlythe potassium salt (acesulfame-K), and sodium and calcium salts thereof.In still another embodiment, the excipient may include a taste-maskingagent. Taste-masking materials include cellulose hydroxypropyl ethers(HPC); low-substituted hydroxypropyl ethers (L-HPC); cellulosehydroxypropyl methyl ethers (HPMC); methylcellulose polymers andmixtures thereof; polyvinyl alcohol (PVA); hydroxyethylcelluloses;carboxymethylcelluloses and salts thereof; polyvinyl alcohol andpolyethylene glycol co-polymers; monoglycerides or triglycerides;polyethylene glycols; acrylic polymers; mixtures of acrylic polymerswith cellulose ethers; cellulose acetate phthalate; and combinationsthereof.

The weight fraction of the excipient or combination of excipients in thecomposition may be about 98% or less, about 95% or less, about 90% orless, about 85% or less, about 80% or less, about 75% or less, about 70%or less, about 65% or less, about 60% or less, about 55% or less, about50% or less, about 45% or less, about 40% or less, about 35% or less,about 30% or less, about 25% or less, about 20% or less, about 15% orless, about 10% or less, about 5% or less, about 2%, or about 1% or lessof the total weight of the composition.

(III) Methods for Treating Conditions Using (+)-Morphinans ComprisingTLR9 Antagonist Activity Either Alone or in Combination with AnotherTherapeutic Agent

Another aspect of the present invention encompasses methods for treatinga condition in a subject in need thereof. In general, the methodcomprises administering to the subject at least one (+)-morphinancomprising TLR9 antagonist activity either alone or in combination withat least one additional therapeutic agent. A variety of disorders ordisease states may be treated with the compounds of the invention.Suitable disorders and diseases that may be treated include, but are notlimited to, pain conditions, inflammatory disorders, acetaminophentoxicity, autoimmune disorders, neurodegenerative disorders, and cancer.

The subject to be treated may be any subject diagnosed as having one ofthe indicated conditions. Moreover, the subject to be treated may be inneed of treatment for the one of the conditions. That is, the subjecthas been diagnosed with the condition or is at risk for developing thecondition, and consequently, is in need of treatment for the condition.The subject may be diagnosed with the condition using diagnostic orclinical tests that are well known. Furthermore, those of skill in theart appreciate that different diagnostic or clinical tests are used todiagnosis the different conditions or disorders. The diagnostic toolsinclude, without limit, physical examination, patient history, screeningtests, laboratory tests, molecular tests, genomic tests, imaging tools,physical tests, mental tests, and the like. Since the perception of painmay be quite subjective, tools such as the McGill Pain Questionnaire maybe used to assess the quality of pain (e.g., sharp, stabbing, squeezing,etc.), and the intensity of pain may be quantified using a numericalscale that ranges from 0 to 10. Skilled diagnosticians are familiar withother indicators of pain.

In general, the subject will be a human. Without departing from thescope of the invention, however, other mammalian subjects may be used.Suitable mammalian subjects include; companion animals, such as cats anddogs; livestock animals, such as cows, pigs, horses, sheep, and goats;zoo animals; and research animals, such as non-human primates androdents.

(a) Conditions

(i) Pain Conditions

In one embodiment, the (+)-morphinan having TLR9 antagonist activity ora pharmaceutically acceptable salt thereof may be used alone or incombination with at least one additional therapeutic agent for thetreatment of a pain condition in a subject. In general, the subject tobe treated has been diagnosed as having a pain condition. As usedherein, the term “pain” refers to the unpleasant sensory and emotionalexperience associated with actual or perceived tissue damage by anoxious stimulus. The pain may be acute or chronic pain. For example,the pain may be traumatic or inflammatory pain, which results frominjury to non-neural tissue. Non-limiting examples of traumatic orinflammatory pain include arachnoiditis, arthritis, back pain, burnpain, central pain syndrome, cancer pain, headaches (includingmigraines, cluster, and tension headaches); head and facial pain, musclepain (including fibromyalgia), myofascial pain syndromes; reflexsympathetic dystrophy syndrome, repetitive stress injuries, sciatica,shingles and other skin disorders, sports injuries, spinal stenosis,surgical pain, temporomandibular disorders, trauma, and/or vasculardisease or injury.

Alternatively, the pain may be neuropathic pain, which results frominjury to or inflammation of the central or peripheral nervous system.Neuropathic pain may occur in any part of the body and is frequentlydescribed as a hot, burning sensation, which can be devastating to theaffected individual. Neuropathic pain may be acute or chronic; it mayresult from diseases that affect nerves (such as diabetes), from trauma,surgical procedures, arthritis, AIDS, burn injuries, cerebral or lumbarspine disease, fibromyalgia, post-eschemic pain, tumors, viralneuralgias, or, because chemotherapy drugs can affect nerves, it may bea consequence of cancer treatment. Among the many neuropathic painconditions are diabetic neuropathy (which results from nerve damagesecondary to vascular problems that occur with diabetes); reflexsympathetic dystrophy syndrome, which may follow injury; phantom limband post-amputation pain, which may result from the surgical removal ofa limb; post-herpetic neuralgia, which may occur after an outbreak ofshingles; and complex regional pain syndrome or central pain syndrome,which may result from trauma to the brain or spinal cord.

Characteristic symptoms of neuropathic pain include hyperesthesia (i.e.,enhanced sensitivity to a natural stimulus); allodynia (i.e., widespreadtenderness or hypersensitivity to tactile stimuli); hyperalgesia (i.e.,abnormal sensitivity to pain); spontaneous burning pain; and/or phantompain (i.e., perception of pain that is non-existent). Hyperesthesiainvolves an unusual increased or altered sensitivity to sensory stimuli,including for example, acoustic, cerebral, gustatory, muscular,olfactory, onelric, optic, or tactile. As an example, a painfulsensation from a normally painless touch stimulus. Allodynia involves anintensified, unpleasant, and painful perception of stimuli triggered byheat or by contact, which is based on a lowering of the pain thresholdfor these stimuli, including, for example, a non-noxious stimulus tonormal skin. Hyperalgesia involves the excessive perception of a varietyof stimuli, again based on a lowering of the pain threshold and thus anabnormally increased pain sense, including for example, auditory ormuscular stimuli. Phantom pain involves a perception of pain in a limbthat is non-existent, such as perceived pain in a limb that has beenamputated, i.e. phantom limb syndrome.

(ii) Inflammatory Disorders

In another embodiment, the (+)-morphinan having TLR9 antagonist activityor a pharmaceutically acceptable salt thereof may be used alone or incombination with at least one additional therapeutic agent for thetreatment of inflammation in a subject. For example, the inflammatorydisorder may be arthritis including, but not limited to, rheumatoidarthritis, spondyloarthropathies, gouty arthritis, osteoarthritis,systemic lupus erythematosus, or juvenile arthritis. In someembodiments, the inflammation may be associated with asthma, allergicrhinitis, sinus diseases, bronchitis, tuberculosis, acute pancreatitis,sepsis, infectious diseases, menstrual cramps, premature labor,tendinitis, bursitis, skin-related conditions such as psoriasis, eczema,atopic dermatitis, urticaria, dermatitis, contact dermatitis, and burns,or from post-operative inflammation including from ophthalmic surgery(e.g., cataract surgery, refractive surgery, etc.) or from organtransplantation surgery (e.g., hepatic transplantation, renaltransplantation, lung transplantation, etc.). In a further embodiment,the inflammatory disorder may be a gastrointestinal condition such asinflammatory bowel disease, Crohn's disease, gastritis, irritable bowelsyndrome, chronic cholecystitis, or ulcerative colitis. In yet anotherembodiment, the inflammation may be associated with diseases such asvascular diseases, migraine headaches, periarteritis nodosa,thyroiditis, aplastic anemia, Hodgkin's disease, sclerodoma, rheumaticfever, type I diabetes, neuromuscular junction disease includingmyasthenia gravis, white matter disease including multiple sclerosis,sarcoidosis, nephrotic syndrome, Behcet's syndrome, polymyositis,gingivitis, nephritis, hypersensitivity, swelling occurring afterinjury, myocardial ischemia, allergic rhinitis, respiratory distresssyndrome, systemic inflammatory response syndrome (SIRS),cancer-associated inflammation, reduction of tumor-associatedangiogenesis, endotoxin shock syndrome, atherosclerosis, and the like.In an alternate embodiment, the inflammatory disorder may be associatedwith an ophthalmic disease, such as retinitis, retinopathies, uveitis,ocular photophobia, or of acute injury to the eye tissue. In stillanother embodiment, the inflammation may be a pulmonary inflammation,such as that associated with viral infections or cystic fibrosis,chronic obstructive pulmonary disease, or acute respiratory distresssyndrome. The inflammatory disorder may also be associated with tissuerejection, graft v. host diseases, delayed-type hypersensitivity, aswell as immune-mediated and inflammatory elements of CNS diseases suchas Alzheimer's, Parkinson's, multiple sclerosis, and the like.

(iii) Acetaminophen Toxicity

In still another embodiment, the (+)-morphinan having TLR9 antagonistactivity or a pharmaceutically acceptable salt thereof may also be aloneor in combination with at least one other therapeutic agent to treatacetaminophen toxicity (also known as paracetamol toxicity). High levelsof acetaminophen may lead to damage of the liver (i.e.,acetaminophen-induced hepatotoxicity) or the kidney (i.e.,acetaminophen-induced nephrotoxicity). Acetaminophen toxicity may resultfrom an acute overdose of acetaminophen or a chronic overdoseacetaminophen. The amount of ingested acetaminophen at which toxicityoccurs may be reduced upon chronic ethanol use, malnourishment, ordiminished nutritional status, fasting, or viral illness withdehydration, or use of certain pharmaceutical agents that interact withthe enzyme systems that metabolize acetaminophen.

Acetaminophen-induced hepatotoxicity may be manifested by cellularoxidative damage, mitochondrial dysfunction, and a subsequentinflammatory response. Cellular damage may be monitored by elevatedlevels of serum alanine transaminase (ALT) or serum aspartatetransaminase (AST), and the inflammatory response may be monitored byincreased levels of pro-interleukin(IL)-1 beta transcript levels.Acetaminophen-induced hepatotoxicity also may lead to hepatocellularinjury, death, and centrilobular (zone III) liver necrosis. Similarenzymatic reactions occur in the kidney, and may contribute to somedegree of extra-hepatic organ dysfunction.

(iv) Autoimmune Disorders

In yet another embodiment, the (+)-morphinan having TLR9 antagonistactivity or a pharmaceutically acceptable salt thereof may also be aloneor in combination with at least one other therapeutic agent to treat anautoimmune disease or disorder. The autoimmune disorder may be systemic,such as Lupus, wherein many tissues or organs are affected or damaged.Alternatively, the autoimmune disorder may be localized, such as type Idiabetes mellitus, wherein a single organ or tissue is damaged oraffected. Non-limiting examples of autoimmune disorders include acutedisseminated encephalomyelitis (ADEM), Addison's disease, alopeciaareata, antiphospholipid antibody syndrome (APS), autoimmune hemolyticanemia, autoimmune hepatitis, autoimmune inner ear disease, bullouspemphigoid, celiac disease, Chagas disease, chronic obstructivepulmonary disease, Crohn's disease, dermatomyositis, diabetes mellitustype 1, endometriosis, Goodpasture's syndrome, Graves' disease,Guillain-Barre syndrome (GBS), Hashimoto's thyroiditis, hidradenitissuppurativa, Kawasaki disease, IgA nephropathy, idiopathicthrombocytopenic purpura, interstitial cystitis, lupus erythematosus(Lupus), mixed connective tissue disease, morphea, multiple sclerosis,myasthenia gravis, narcolepsy, neuromyotonia, pemphigus vulgaris,pernicious anemia, psoriasis, psoriatic arthritis, polymyalgiarheumatica, polymyositis, primary biliary cirrhosis, rheumatoidarthritis and juvenile rheumatoid arthritis, schizophrenia,schleroderma, sclerosing cholangitis, Sjogren's syndrome, stiff personsyndrome, temporal arteritis/giant cell arteritis, ulcerative colitis,vasculitis, vitiligo, and Wegener's granulomatosis.

(v) Neurodegenerative Disorders

In an alternate embodiment, the (+)-morphinan having TLR9 antagonistactivity or a pharmaceutically acceptable salt thereof may be used aloneor in combination with at least one other therapeutic agent to treat aneurodegenerative disorder. Non-limiting examples of neurodegenerativedisorders include adrenal leukodystrophy, aging-related disorders anddementias, alcoholism, Alexander's disease, Alper's disease, Alzheimer'sdisease, amyotrophic lateral sclerosis (Lou Gehrig's Disease), ataxiatelangiectasia, Batten disease (also known asSpielmeyer-Vogt-Sjögren-Batten disease), bovine spongiformencephalopathy (BSE), canavan disease, cerebral palsy, Cockaynesyndrome, corticobasal degeneration (CBD), Creutzfeldt-Jakob disease,familial fatal insomnia, frontotemporal lobar degeneration, frontaltemporal dementias (FTDs), Huntington's disease, HIV-associateddementia, Kennedy's disease, Krabbe's disease, Lewy body disease,neuroborreliosis, Machado-Joseph disease (spinocerebellar ataxia type3), multiple system atrophy, multiple sclerosis, narcolepsy, NiemannPick disease, Parkinson disease, Pelizaeus-Merzbacher disease, Pick'sdisease, primary lateral sclerosis, progressive supranuclear palsy(PSP), psychotic disorders, Refsum's disease, Sandhoff disease,Schilder's disease, schizoaffective disorder, schizophrenia, stroke,subacute combined degeneration of spinal cord secondary to perniciousanemia, spinocerebellar ataxia, spinal muscular atrophy,Steele-Richardson-Olszewski disease, Tabes dorsalis, and toxicencephalopathy.

(vi) Cancers

In still another embodiment, the (+)-morphinan having TLR9 antagonistactivity or a pharmaceutically acceptable salt thereof may be used incombination with a chemotherapeutic agent to treat a neoplasm or acancer. The neoplasm may be malignant or benign, the cancer may beprimary or metastatic; the neoplasm or cancer may be early stage or latestage. Non-limiting examples of neoplasms or cancers that may be treatedinclude acute lymphoblastic leukemia, acute myeloid leukemia,adrenocortical carcinoma, AIDS-related cancers, AIDS-related lymphoma,anal cancer, appendix cancer, astrocytomas (childhood cerebellar orcerebral), basal cell carcinoma, bile duct cancer, bladder cancer, bonecancer, brainstem glioma, brain tumors (cerebellar astrocytoma, cerebralastrocytoma/malignant glioma, ependymoma, medulloblastoma,supratentorial primitive neuroectodermal tumors, visual pathway andhypothalamic gliomas), breast cancer, bronchial adenomas/carcinoids,Burkitt lymphoma, carcinoid tumors (childhood, gastrointestinal),carcinoma of unknown primary, central nervous system lymphoma (primary),cerebellar astrocytoma, cerebral astrocytoma/malignant glioma, cervicalcancer, childhood cancers, chronic lymphocytic leukemia, chronicmyelogenous leukemia, chronic myeloproliferative disorders, coloncancer, cutaneous T-cell lymphoma, desmoplastic small round cell tumor,endometrial cancer, ependymoma, esophageal cancer, Ewing's sarcoma inthe Ewing family of tumors, extracranial germ cell tumor (childhood),extragonadal germ cell tumor, extrahepatic bile duct cancer, eye cancers(intraocular melanoma, retinoblastoma), gallbladder cancer, gastric(stomach) cancer, gastrointestinal carcinoid tumor, gastrointestinalstromal tumor, germ cell tumors (childhood extracranial, extragonadal,ovarian), gestational trophoblastic tumor, gliomas (adult, childhoodbrain stem, childhood cerebral astrocytoma, childhood visual pathway andhypothalamic), gastric carcinoid, hairy cell leukemia, head and neckcancer, hepatocellular (liver) cancer, Hodgkin lymphoma, hypopharyngealcancer, hypothalamic and visual pathway glioma (childhood), intraocularmelanoma, islet cell carcinoma, Kaposi sarcoma, kidney cancer (renalcell cancer), laryngeal cancer, leukemias (acute lymphoblastic, acutemyeloid, chronic lymphocytic, chronic myelogenous, hairy cell), lip andoral cavity cancer, liver cancer (primary), lung cancers (non-smallcell, small cell), lymphomas (AIDS-related, Burkitt, cutaneous T-cell,Hodgkin, non-Hodgkin, primary central nervous system), macroglobulinemia(Waldenström), malignant fibrous histiocytoma of bone/osteosarcoma,medulloblastoma (childhood), melanoma, intraocular melanoma, Merkel cellcarcinoma, mesotheliomas (adult malignant, childhood), metastaticsquamous neck cancer with occult primary, mouth cancer, multipleendocrine neoplasia syndrome (childhood), multiple myeloma/plasma cellneoplasm, mycosis fungoides, myelodysplastic syndromes,myelodysplastic/myeloproliferative diseases, myelogenous leukemia(chronic), myeloid leukemias (adult acute, childhood acute), multiplemyeloma, myeloproliferative disorders (chronic), nasal cavity andparanasal sinus cancer, nasopharyngeal carcinoma, neuroblastoma,non-Hodgkin lymphoma, non-small cell lung cancer, oral cancer,oropharyngeal cancer, osteosarcoma/malignant fibrous histiocytoma ofbone, ovarian cancer, ovarian epithelial cancer (surfaceepithelial-stromal tumor), ovarian germ cell tumor, ovarian lowmalignant potential tumor, pancreatic cancer, pancreatic cancer (isletcell), paranasal sinus and nasal cavity cancer, parathyroid cancer,penile cancer, pharyngeal cancer, pheochromocytoma, pineal astrocytoma,pineal germinoma, pineoblastoma and supratentorial primitiveneuroectodermal tumors (childhood), pituitary adenoma, plasma cellneoplasia, pleuropulmonary blastoma, primary central nervous systemlymphoma, prostate cancer, rectal cancer, renal cell carcinoma (kidneycancer), renal pelvis and ureter transitional cell cancer,retinoblastoma, rhabdomyosarcoma (childhood), salivary gland cancer,sarcoma (Ewing family of tumors, Kaposi, soft tissue, uterine), Sezarysyndrome, skin cancers (nonmelanoma, melanoma), skin carcinoma (Merkelcell), small cell lung cancer, small intestine cancer, soft tissuesarcoma, squamous cell carcinoma, squamous neck cancer with occultprimary (metastatic), stomach cancer, supratentorial primitiveneuroectodermal tumor (childhood), T-Cell lymphoma (cutaneous),testicular cancer, throat cancer, thymoma (childhood), thymoma andthymic carcinoma, thyroid cancer, thyroid cancer (childhood),transitional cell cancer of the renal pelvis and ureter, trophoblastictumor (gestational), unknown primary site (adult, childhood), ureter andrenal pelvis transitional cell cancer, urethral cancer, uterine cancer(endometrial), uterine sarcoma, vaginal cancer, visual pathway andhypothalamic glioma (childhood), vulvar cancer, Waldenströmmacroglobulinemia, and Wilms tumor (childhood).

(vii) Hepatic Disorders

In an alternate embodiment, the (+)-morphinans having TLR9 antagonistactivity or a pharmaceutically acceptable salt thereof may be used aloneor in combination with at least one other therapeutic agent to treathepatic disorders. As detailed above in sections (III)(a)(ii) and(III)(a)(iii), the (+)-morphinans having TLR9 antagonist activity may beused to treat inflammatory disorders and inflammatory-mediatedacetaminophen hepatotoxicity. Accordingly, the (+)-morphinans havingTLR9 antagonist activity may be used to treat inflammatory disorders ofthe liver.

Inflammatory disorders of the liver include ischemia-reperfusion injury,which is an inflammation-mediated injury. Ischemia-reperfusion injury isan important cause of liver damage occurring during surgical proceduresincluding hepatic resection and liver transplantation, and representsthe main underlying cause of graft dysfunction post-transplantation.Liver transplantation is a viable treatment option for any acute orchronic condition resulting in irreversible liver dysfunction. Acutehepatic injuries include, without limit, drug-induced liver failure(e.g., acetaminophen overdose), acute viral hepatitis, ischemichepatitis, and acute hepatic necrosis. Chronic hepatic conditionsleading to liver dysfunction include long-term alcohol abuse, long-termuntreated hepatitis C infection, long-term untreated hepatitis Binfection, cirrhosis, hemochromatosis, Wilson's disease, autoimmunediseases, the like. The (+)-morphinans having TLR9 antagonist activitymay reduce or prevent ischemia-reperfusion injury during hepaticresections and/or liver transplantations of normal or steatotic (i.e.,fatty) livers.

In an exemplary iteration of this embodiment, R is methylcyclopropyl,R^(a) is hydrogen, R^(b) is methyl, and Y is hydroxy. In anotheriteration, R is methyl, R^(a) is methyl, R^(b) ismethylacetylmethylester; and Y is hydrogen. In yet another iteration, Ris allyl, R^(a) is methyl, R^(b) is propyl; and Y is hydrogen. In afurther iteration, R is allyl, R^(a) is methyl, R^(b) is ethylalcohol;and Y is hydrogen.

Other inflammatory disorders of the liver for which treatment with the(+)-morphinans having TLR9 antagonist activity may be indicated include,without limit, hepatic steatosis (also called steatohepatitis or fattyliver disease), hepatic steatosis post-liver transplantation, hepatitis,autoimmune hepatitis, alcoholic hepatitis, viral hepatitis,non-alcoholic fatty liver disease (also called non-alcoholicsteatohepatitis), ischemic hepatitis, metabolic disorder hepatitis,chronic liver inflammation, hepatic fibrosis, cholestasis, steatosis,hepatic granulomas, acute or chronic liver transplant rejection andmetabolic conditions, hepatocellular carcinoma, cholangiocarcinoma,primary sclerosing cholangitis, cirrhosis, primary biliary cirrhosis,zonal necrosis, hemochromatosis, Wilson's disease, alpha 1-antitrypsindeficiency, glycogen storage disease type II, Gilbert's syndrome, portalhypertension, portal vein thrombosis, ascites, variceal bleeding, buddchiari, hepatic viral infections, hepatic bacterial infections, hepaticparasitic infections, and liver abscesses.

(b) Treatment Formulations

(i) Formulations Comprising at Least One (+)-Morphinan Having TLR9Antagonist Activity

In some embodiments, the method comprises administering to the subjectat least one (+)-morphinan compound having TLR9 antagonist activity or apharmaceutically acceptable salt thereof.

In one embodiment, the (+)-morphinan administered to the subject may bea compound comprising Formula (IIa) or a pharmaceutically acceptablesalt thereof:

wherein:

-   -   R is chosen from hydrogen, methyl, alkyl, alkenyl, allyl,        methylcycloalkyl, methylcyclopropyl, methylcyclobutyl,        methylaryl, methylphenyl, acyl, acylalkyl, acylcycloalkyl,        acylcyclopropyl, acylcyclobutyl, acylaryl, acylphenyl, acyloxy,        acyloxyalkyl, acyloxyaryl, acyloxyphenyl, alkoxy, and        alkoxyalkyl;    -   R¹ is chosen from hydrogen, halo, hydroxy, alkoxy, alkoxyalkyl,        alkoxyalkenyl, aryloxy, alkyl, alkenyl, aryl, heteroaryl, and        amine;    -   R³ is chosen from hydroxy, alkoxy, aryloxy, acyl, acyloxy, and        protected hydroxy;    -   A₁ is chosen from hydrogen, hydroxy, keto, alkoxy, acyl,        acyloxy, amino, amide, alkylamine, hydroxyalkylamine,        carboxylalkylamine, alkylcarboxylalkylamine, arylamine, alkyl,        alkenyl, aryl, substituted alkyl, substituted alkenyl, and        substituted aryl; provided, however, that when A₁ is keto, R¹ is        other than hydrogen;    -   Y is chosen from hydrogen, hydroxy, alkoxy, and protected        hydroxy; and    -   the dashed line represents an optional double bond.

In one preferred embodiment, the (+)-morphinan administered to thesubject may be a compound comprising Formula (IIb) or a pharmaceuticallyacceptable salt thereof:

wherein:

-   -   R is chosen from hydrogen, alkyl, methyl, allyl,        methylcyclopropyl, and methylcyclobutyl;    -   R^(a) is chosen from hydrogen, methyl, alkyl, phenyl, and        benzyl;    -   R^(b) is chosen from hydrogen, methyl, alkyl, alkylalcohol,        alkylcarboxyl, alkylcarboxylalkylester, acyl, acylalkyl,        acylaryl, acyloxy, acyloxyalkyl, acyloxyaryl; and    -   Y is chosen from hydrogen and hydroxy.

In an exemplary iteration of this embodiment, R is methylcyclopropyl,R^(a) is hydrogen, R^(b) is methyl, and Y is hydroxy. In anotheriteration, R is methyl, R^(a) is methyl, R^(b) ismethylacetylmethylester; and Y is hydrogen. In yet another iteration, Ris allyl, R^(a) is methyl, R^(b) is propyl; and Y is hydrogen. In afurther iteration, R is allyl, R^(a) is methyl, R^(b) is ethylalcohol;and Y is hydrogen.

In another preferred embodiment, the (+)-morphinan administered to thesubject may be a compound comprising Formula (IIc) or a pharmaceuticallyacceptable salt thereof:

wherein:

-   -   R is chosen from hydrogen, methyl, alkyl, allyl,        methylcyclopropyl, and methylcyclobutyl;    -   R¹ is chosen from hydrogen, halo, hydroxy, alkoxy, amine, aryl,        heteroaryl, and furanyl;    -   R^(a) is chosen from hydrogen, methyl, alkyl, phenyl, and        benzyl;    -   R^(c) is chosen from hydroxy, methoxy, alkoxy, keto, and acyl;        provided, however, that when R^(c) is keto, R¹ is not hydrogen;    -   Y is chosen from hydrogen and hydroxy; and    -   the dashed line represents an optional double bond.

In one iteration of this embodiment, R is methylcyclopropyl, R¹ ishydrogen, R^(a) is methyl, R^(c) is hydroxy; and Y is hydrogen. Inanother iteration, R is methylcyclopropyl, R¹ is hydrogen, R^(a) ismethyl, R^(c) is methoxy, Y is hydrogen, and the optional double bond ispresent. In a further iteration, R is allyl, R¹ is furanyl, R^(a) ismethyl, R^(c) is keto, and Y is hydrogen. In still another iteration, Ris hydrogen, R¹ is hydrogen, R^(a) is methyl, R^(c) is methoxy, Y ishydrogen, and the optional double bond is present.

In another embodiment, the (+)-morphinan administered to the subject maybe a compound comprising Formula (IIIa) or a pharmaceutically acceptablesalt thereof:

wherein:

-   -   R is chosen from hydrogen, methyl, alkyl, alkenyl, allyl,        methylcycloalkyl, methylcyclopropyl, methylcyclobutyl,        methylaryl, methylphenyl, acyl, acylalkyl, acylcycloalkyl,        acylcyclopropyl, acylcyclobutyl, acylaryl, acylphenyl, acyloxy,        acyloxyalkyl, acyloxyaryl, acyloxyphenyl, alkoxy, and        alkoxyalkyl; and    -   R³, R⁴, and R⁷ are independently chosen from hydrogen, hydroxy,        alkoxy, aryloxy, acyloxy, amine, halo, and protected hydroxy.

In one iteration of this embodiment, R is allyl, R³ is methoxy, R⁴ ishydroxy, and R⁷ is methoxy.

(ii) Combination Formulations

In other embodiments, the treatment method comprises administering tothe subject a combination formulation comprising at least one(+)-morphinan TLR9 antagonist, as disclosed herein, and at least oneother therapeutic agent. It is envisioned that when the combinationformulation comprises more than one additional therapeutic agent, thedifferent agents may be drawn from one of the classes listed below, orthe different agents may be drawn from different classes listed below.

In one embodiment, the combination formulation may comprise a(+)-morphinan or a pharmaceutically acceptable salt thereof and acompound comprising Formula (IV) or a pharmaceutically acceptable saltthereof, as detailed above in section (II).

In another embodiment, the combination formulation may comprise a(+)-morphinan or a pharmaceutically acceptable salt thereof, as detailedabove in sections (I)(a), (I)(b), and (I)(c), and an analgesic agent.The analgesic may be an (−)-opioid analgesic. Alternatively, theanalgesic may be a non-opioid analgesic. Non-limiting examples ofsuitable opioid analgesics include buprenorphine, butorphanol, codeine,dihydrocodeine, dihydromorphine, etorphine, fentanyl, hydrocodone,hydromorphone, levophanol, meperidine, methadone, morphine, nalbuphine,norcodeine, normorphine, oxycodone, oxymorphone, pentazocine, andpropoxyphene. In some combinations comprising an opioid analgesic, theconcentration or dose of the opioid analgesic in the combinationformulation may be sub-analgesic. Examples of suitable non-opioidanalgesics include without limit acetylsalicylic acid, acetaminophen(paracetamol), ibuprofen, ketoprofen, indomethacin, diflunisol,naproxen, ketorolac, dichlophenac, tolmetin, sulindac, phenacetin,piroxicam, and mefamanic acid. In further embodiments, the analgesic maycomprise a combination of an opiate analgesic and a non-opioidanalgesic. For example, acetaminophen may be combined with codeine,hydrocodone, oxycodone, propoxyphene, or another opioid analgesic. In anexemplary embodiment, the combination may comprise a (+)-morphinancomprising Formulas (I), (II), (IIa), (IIb), (IIc), (III), or (IIIa),and acetaminophen. The concentration of acetaminophen in such acombination may be lower than in currently available acetaminophencombination formulations.

In a further embodiment, the combination formulation may comprise a(+)-morphinan or a pharmaceutically acceptable salt thereof, as detailedabove in sections (I)(a), (I)(b), and (I)(c), and an anti-inflammatoryagent. The anti-inflammatory agent may be a glucocorticoid steroid suchas the naturally occurring hydrocortisone (cortisol), or syntheticglucocorticoids such as prednisone, prednisolone, methylprednisolone,dexamethasone, betamethasone, triamcinolone, beclometasone,fludrocortisones, deoxycorticosterone, alclometasone, fluocinonide,aldosterone, and derivatives thereof. Alternatively, theanti-inflammatory agent may be a non-steroidal anti-inflammatory agent(NSAID). Non-limiting examples of suitable NSAIDs includeacetylsalicylic acid (aspirin), celecoxib, choline magnesium salicylate,Cox-2 inhibitors, diclofenac, diflunisal, etodolac, fenoprofen,flufenisal, flurbiprofen, ibuprofen, indomethacin, ketoprofen,ketorolac, meclofenamate, mefenamate, nabumetone, naproxen, oxaprozin,phenylbutazone, piroxicam, salsalate, sulindac, tolmetin, valdecoxib,and zomepirac.

In yet another embodiment, the combination formulation may comprise a(+)-morphinan or a pharmaceutically acceptable salt thereof, as detailedabove in sections (I)(a), (I)(b), and (I)(c), and an antibiotic agent.Non-limiting examples of suitable antibiotic agents includeaminoglycosides such as, e.g., amikacin, gentamicin, kanamycin,neomycin, netilmicin, streptomycin, and tobramycin; a carbecephem suchas loracarbef; carbapenems such as, e.g., certapenem, imipenem, andmeropenem; cephalosporins such as, e.g., cefadroxil cefazolin,cephalexin, cefaclor, cefamandole, cephalexin, cefoxitin, cefprozil,cefuroxime, cefixime, cefdinir, cefditoren, cefoperazone, cefotaxime,cefpodoxi me, ceftazidime, ceftibuten, ceftizoxime, and ceftriaxone;macrolides such as, e.g., azithromycin, clarithromycin, dirithromycin,erythromycin, and troleandomycin; monobactam; penicillins such as, e.g.,amoxicillin, ampicillin, carbenicillin, cloxacillin, dicloxacillin,nafcillin, oxacillin, penicillin G, penicillin V, piperacillin, andticarcillin; polypeptides such as, e.g., bacitracin, colistin, andpolymyxin B; quinolones such as, e.g., ciprofloxacin, enoxacin,gatifloxacin, levofloxacin, lomefloxacin, moxifloxacin, norfloxacin,ofloxacin, and trovafloxacin; sulfonamides such as, e.g., mafenide,sulfacetamide, sulfamethizole, sulfasalazine, sulfisoxazole, andtrimethoprim-sulfamethoxazole; tetracyclines such as, e.g.,demeclocycline, doxycycline, minocycline, and oxytetracycline); and anantimicrobial agent such as, e.g., ketoconazole, amoxicillin,cephalexin, miconazole, econazole, acyclovir, and nelfinavir.

In another alternate embodiment, the combination formulation maycomprise a (+)-morphinan or a pharmaceutically acceptable salt thereof,as detailed above in sections (I)(a), (I)(b), and (I)(c), and an agentused to treat acetaminophen toxicity. Suitable agents includeacetylcysteine (also called N-acetylcysteine), glutathione, andactivated charcoal.

In still another embodiment, the combination formulation may comprise a(+)-morphinan or a pharmaceutically acceptable salt thereof, as detailedabove in sections (I)(a), (I)(b), and (I)(c), and an autoimmunetherapeutic agent. Non-limiting examples of suitable autoimmunetherapeutic agents include immunosuppressants such as azathioprine,chlorambucil, cyclophosphamide, cyclosporine, mycophenolate, ormethotrexate; corticosteroids such as prednisone; the psoriasistreatment agent alefacept; TNF blockers such as etanercept, infliximab,or adalimumab; white blood cell blockers such as abatacept or ritaximab;the leprosy drug clofazimine; and chemotherapeutic agents such asvorinostat.

In a further embodiment, the combination formulation may comprise a(+)-morphinan or a pharmaceutically acceptable salt thereof, as detailedabove in sections (I)(a), (I)(b), and (I)(c), and a neurodegenerativedisorder therapeutic agent. Typically, the neurodegenerative disordertherapeutic agent is tailored to the specific neurodegenerative disorderto be treated. Suitable therapeutic agents for the treatment ofParkinson disease include, without limit, levadopa (i.e., L-DOPA); adecarboxylase inhibitor such as carbidopa; a direct acting dopamineagonist such bromocriptine, pergolide, ropinirole or pramipexole; adopamine uptake inhibitor such as amantadine; an anticholinergic such astrihexyphenidyl or benztropine mesylate; a monoamine oxidase B inhibitorsuch as L-deprenyl; a catechol-O-methyltranferase inhibitor such astolcapone; spheramine; and combinations thereof. Non-limiting examplesof suitable therapeutic agents for the treatment of Alzheimer's diseaseinclude cholinesterase inhibitors such as donepezil, rivastigmine,galantamine, and the like; NMDA receptor antagonists such as memantine;and Alzheimer's specific agents such as tramiprosate, tarenflubil,phenserine, and the like. Targeted therapeutic agents used to treatHuntington's disease include, without limit, tetrabenazine, xenazine,and so forth. Non-limiting examples of therapeutic agents targeted totreat amyotrophic lateral sclerosis (ALS) include riluzole, mecaserminrinfabate, and the like.

In an alternate embodiment, the combination formulation may comprise a(+)-morphinan or a pharmaceutically acceptable salt thereof, as detailedabove in sections (I)(a), (I)(b), and (I)(c), and a chemotherapeuticagent. The chemotherapeutic agent may be a cytotoxic agent that affectsrapidly dividing cells in general, or it may be a targeted therapeuticagent that affects the deregulated proteins of cancer cells. Forexample, the chemotherapeutic agent may be an alkylating agent, ananti-metabolite, an anti-tumor antibiotic, an anti-cytoskeletal agent, atopoisomerase inhibitor, an anti-hormonal agent, a targeted therapeuticagent, or a combination thereof. Non-limiting examples of alkylatingagents include altretamine, benzodopa, busulfan, carboplatin,carboquone, carmustine, chlorambucil, chlornaphazine, cholophosphamide,chlorozotocin, cisplatin, cyclosphosphamide, dacarbazine (DTIC),estramustine, fotemustine, ifosfamide, improsulfan, lomustine,mechlorethamine, mechlorethamine oxide hydrochloride, melphalan,meturedopa, nimustine, novembichin, phenesterine, piposulfan,prednimustine, ranimustine; temozolomide, thiotepa, triethylenemelamine,trietylenephosphoramide, triethylenethiophosphaoramide,trimethylolomelamine, trofosfamide, uracil mustard and uredopa. Suitableanti-metabolites include, but are not limited to aminopterin,ancitabine, azacitidine, 6-azauridine, capecitabine, carmofur,cytarabine or cytosine arabinoside (Ara-C), dideoxyuridine, denopterin,doxifluridine, enocitabine, floxuridine, fludarabine, 5-fluorouracil(5-FU), gemcetabine, leucovorin (folinic acid), 6-mercaptopurine,methotrexate, pemetrexed, pteropterin, thiamiprine, trimetrexate, andthioguanine. Non-limiting examples of suitable anti-tumor antibioticsinclude aclacinomysin, actinomycin, adriamycin, authramycin, azaserine,bleomycins, cactinomycin, calicheamicin, carabicin, caminomycin,carzinophilin, chromomycins, dactinomycin, daunorubicin, detorubicin,6-diazo-5-oxo-L-norleucine, doxorubicin, epirubicin, esorubicin,idarubicin, marcellomycin, mitomycins, mycophenolic acid, nogalamycin,olivomycins, peplomycin, potfiromycin, puromycin, quelamycin,rodorubicin, streptonigrin, streptozocin, tubercidin, ubenimex,zinostatin, and zorubicin. Non-limiting examples of suitableanti-cytoskeletal agents include colchicines, docetaxel, macromycin,paclitaxel (taxol), vinblastine, vincristine, vindesine, andvinorelbine. Suitable topoisomerase inhibitors include, but are notlimited to, amsacrine, etoposide (VP-16), irinotecan, RFS 2000,teniposide, and topotecan. Non-limiting examples of suitableanti-hormonal agents such as aminoglutethimide, aromatase inhibiting4(5)-imidazoles, bicalutamide, finasteride, flutamide, goserelin,4-hydroxytamoxifen, keoxifene, leuprolide, LY117018, mitotane,nilutamide, onapristone, raloxifene, tamoxifen, toremifene, andtrilostane. Non-limiting examples of targeted therapeutic agents includea monoclonal antibody such as alemtuzumab, bevacizumab, capecitabine,cetuximab, gemtuzumab, heregulin, rituximab, trastuzumab; a tyrosinekinase inhibitor such as imatinib mesylate; and a growth inhibitorypolypeptide such as erythropoietin, interleukins (e.g., IL-1, IL-2,IL-3, IL-6), leukemia inhibitory factor, interferons, thrombopoietin,TNF-α, CD30 ligand, 4-1 BB ligand, and Apo-1 ligand.

Those of skill in the art appreciate that pharmaceutically acceptablesalts, acids, or derivatives of any of the above listed agents may beincluded in the combination formulations.

(c) Dosage Forms

The formulations detailed above in section (III)(b) may be administeredby a number of different means that will deliver a therapeuticallyeffective dose. For example, the formulations may be administeredorally, parenterally, by inhalation spray, rectally, intradermally,intrathecally, transdermally, or topically in dosage unit formulationscontaining conventional nontoxic pharmaceutically acceptable carriers,adjuvants, and vehicles as desired. Topical administration may alsoinvolve the use of transdermal administration such as transdermalpatches or iontophoresis devices. Formulation of therapeutic agents isdiscussed in, for example, Gennaro, A. R., Remington's PharmaceuticalSciences, Mack Publishing Co., Easton, Pa. (18th ed, 1995), andLiberman, H. A. and Lachman, L., Eds., Pharmaceutical Dosage Forms,Marcel Dekker Inc., New York, N.Y. (1980).

Preparations for oral administration generally contain inertpharmaceutically acceptable excipients in addition to the activepharmaceutical ingredient. Oral preparations may be enclosed in gelatincapsules or compressed into tablets. Common excipients used in suchpreparations include pharmaceutically compatible fillers/diluents suchas microcrystalline cellulose, hydroxypropyl methylcellulose, starch,lactose, sucrose, glucose, mannitol, sorbitol, dibasic calciumphosphate, or calcium carbonate; binding agents such as alginic acid,carboxymethylcellulose, microcrystalline cellulose, gelatin, gumtragacanth, or polyvinylpyrrolidone; disintegrating agents such asalginic acid, cellulose, starch, or polyvinylpyrrolidone; lubricantssuch as calcium stearate, magnesium stearate, talc, silica, or sodiumstearyl fumarate; glidants such as colloidal silicon dioxide; sweeteningagents such as sucrose or saccharin; flavoring agents such aspeppermint, methyl salicylate, or citrus flavoring; coloring agents; andpreservatives such as antioxidants (e.g., vitamin A, vitamin C, vitaminE, or retinyl palmitate), citric acid, or sodium citrate. Oralpreparations may also be administered as aqueous suspensions, elixirs,or syrups. For these, the active ingredient may be combined with varioussweetening or flavoring agents, coloring agents, and, if so desired,emulsifying and/or suspending agents, as well as diluents such as water,ethanol, glycerin, and combinations thereof.

For parenteral administration (including subcutaneous, intradermal,intravenous, intramuscular, and intraperitoneal), the preparation may bean aqueous or an oil-based solution. Aqueous solutions may include asterile diluent such as water, saline solution, a pharmaceuticallyacceptable polyol such as glycerol, propylene glycol, or other syntheticsolvents; an antibacterial and/or antifungal agent such as benzylalcohol, methyl paraben, chlorobutanol, phenol, thimerosal, and thelike; an antioxidant such as ascorbic acid or sodium bisulfite; achelating agent such as etheylenediaminetetraacetic acid; a buffer suchas acetate, citrate, or phosphate; and/or an agent for the adjustment oftonicity such as sodium chloride, dextrose, or a polyalcohol such asmannitol or sorbitol. The pH of the aqueous solution may be adjustedwith acids or bases such as hydrochloric acid or sodium hydroxide.Oil-based solutions or suspensions may further comprise sesame, peanut,olive oil, or mineral oil.

For topical (e.g., transdermal or transmucosal) administration,penetrants appropriate to the barrier to be permeated are generallyincluded in the preparation. Transmucosal administration may beaccomplished through the use of nasal sprays, aerosol sprays, tablets,or suppositories, and transdermal administration may be via ointments,salves, gels, patches, or creams as generally known in the art.

The amount of agent that is administered to the subject can and willvary depending upon the type of agent, the subject, the condition beingtreated, and the particular mode of administration. Those skilled in theart will appreciate that dosages may also be determined with guidancefrom Goodman & Goldman's The Pharmacological Basis of Therapeutics,Tenth Edition (2001), Appendix II, pp. 475-493, and the Physicians' DeskReference.

(d) Exemplary Treatment Methods

In exemplary embodiments, the condition to be treated may be a paincondition. The pain condition may be acute pain, traumatic pain, chronicpain, neuropathic pain, or combinations thereof. Generally, the subjectto be treated is diagnosed as having a pain condition and is need oftreatment for the pain condition. In one iteration of this embodiment,the method comprises administering to the subject in need thereof aformulation comprising at least one (+)-morphinan as detailed above insection (III)(b)(i). In another iteration of this embodiment, the methodcomprises administering to the subject in need thereof a formulationcomprising a (+)-morphinan and a compound comprising Formula (IV) asdetailed above in sections (II) and (III)(b)(ii).

(IV) Methods for Inhibiting TLR9 Activation

A further aspect of the present invention provides methods forinhibiting the activation of TLR9. In general, the method comprisescontacting a cell expressing TLR9 with a (+)-morphinan detailed above insections (I)(a)-(c) or a pharmaceutically acceptable salt thereof.

The method of inhibiting the activation of TLR9 may be conducted in vivoor it may be conducted in vitro. Accordingly, the cell expressing TLR9may be disposed in a subject as detailed above. In preferredembodiments, the cell may be a glial cell, a microglial cell, or anastrocyte. In an exemplary embodiment, the cell may be a glial cell inthe central nervous system.

The present invention also provides a method for identifying a(+)-morphinan comprising any of the formulas disclosed herein that maybe therapeutically effective for treating conditions associated withpain and inflammation. Suitable conditions include traumatic orneuropathic pain, an inflammatory disorder, acetaminophen toxicity, anautoimmune disorder, a neurodegenerative disorder, and cancer. Themethod comprises determining whether the compound inhibits TLR9activation. To determine whether the (+)-morphinan inhibits TLR9activation, the method comprises contacting a cell expressing TLR9 withan activation ligand and the (+)-morphinan of interest, wherein TLR9activation is reduced in the presence of the (+)-morphinan as comparedto a control condition in which the cell is contacted with only theactivation ligand. Typically, the cell expressing TLR9 that is contactedwith the (+)-morphinan of interest is in vitro.

Typically, the cell expressing TLR9 will be from a stable cell line.Non-limiting examples of suitable parental cells include HEK293, CHO,BHK, NSO, HDMEC, NHEK, and NHDF cells. In an exemplary embodiment, thecell line may be HEK293. The cells may be engineered to express TLR9using standard procedures well known to those of skill in the art. TLR9may be of mammalian origin, preferably of human origin.

The activation ligand used to activate TLR9 may be methylated DNA,unmethylated DNA, a CpG oligodeoxynucleotide, or anoligodeoxynucleotide. In an exemplary embodiment, the activation ligandmay be CpG oligodeoxynucleotide (ODN) 2006.

The activation of TLR9 in the cell of the in vitro assay may bemonitored by measuring the activity of a reporter, wherein the activityof the reporter is coupled to activation of an adaptor protein or kinasethat mediates TLR9 signaling by producing intracellular signalingmolecules or inducers such as NF-κB or IRF3. Non-limiting examples ofsuitable reporters include luciferase, alkaline phosphatase, and GFP orother fluorescent proteins. The activation of the reporter may bemonitored via luminescence, fluorescence, absorbance, or opticaldensity. In an exemplary embodiment, the reporter is secreted alkalinephosphatase (SEAP) that is induced by NF-κB, and activation of SEAP ismonitored spectrophotometically.

In general, the (+)-morphinan comprising TLR9 antagonist activity mayreduce the activation of TLR9 by at least about 10%. In variousembodiments, the (+)-morphinan may reduce the activation of TLR9 fromabout 10% to about 15%, from about 15% to about 20%, from about 20% toabout 25%, from about 25% to about 30%, from about 30% to about 35%,from about 35% to about 40%, from about 40% to about 45%, from about 45%to about 50%, from about 50% to about 55%, from about 55% to about 60%,from about 60% to about 70%, from about 70% to about 80%, from about 80%to about 90%, or from about 90% to about 99%.

The in vitro screening assay may also be used to determine the optimalinhibitory concentration (or IC₅₀) of a (+)-morphinan comprising TLR9antagonist activity. That is, a dose-response curve may be generated inwhich the concentration of the (+)-morphinan comprising TLR9 antagonistactivity is varied such that the optimal inhibitory concentration may bedetermined.

Definitions

The compounds described herein have asymmetric centers. Compounds of thepresent invention containing an asymmetrically substituted atom may beisolated in optically active or racemic form. All chiral,diastereomeric, racemic forms and all geometric isomeric forms of astructure are intended, unless the specific stereochemistry or isomericform is specifically indicated.

The term “acyl,” as used herein alone or as part of another group,denotes the moiety formed by removal of the hydroxy group from the groupCOOH of an organic carboxylic acid, e.g., RC(O)—, wherein R is R¹, R¹O—,R¹R²N—, or R¹S—, R¹ is hydrocarbyl, heterosubstituted hydrocarbyl, orheterocyclo, and R² is hydrogen, hydrocarbyl, or substitutedhydrocarbyl.

The term “acyloxy,” as used herein alone or as part of another group,denotes an acyl group as described above bonded through an oxygenlinkage (O), e.g., RC(O)O— wherein R is as defined in connection withthe term “acyl.”

The term “alkyl” as used herein describes groups which are preferablylower alkyl containing from one to eight carbon atoms in the principalchain and up to 20 carbon atoms. They may be straight or branched chainor cyclic and include methyl, ethyl, propyl, isopropyl, butyl, hexyl andthe like. A “C₂-C₆ alkyl” refers to an alkyl group containing from twoto six carbon atoms in the principal chain.

The term “alkenyl” as used herein describes groups which are preferablylower alkenyl containing from two to eight carbon atoms in the principalchain and up to 20 carbon atoms. They may be straight or branched chainor cyclic and include ethenyl, propenyl, isopropenyl, butenyl,isobutenyl, hexenyl, and the like. A methylene group is not an alkenylgroup.

As used herein, the term “alkoxy” refers to an alkyl group singularbonded to oxygen, i.e., R—O. Alkyloxy groups include methoxy, ethoxy,propoxy, butoxy, and so forth. The term “C₂-C₆ alkyoxy” describes alkoxygroups having from two to six carbons in the principal chain (i.e.,ethoxy, propoxy, butoxy, and the like).

The term “alkynyl” as used herein describes groups which are preferablylower alkynyl containing from two to eight carbon atoms in the principalchain and up to 20 carbon atoms. They may be straight or branched chainand include ethynyl, propynyl, butynyl, isobutynyl, hexynyl, and thelike.

The term “aromatic” as used herein alone or as part of another groupdenotes optionally substituted homo- or heterocyclic conjugated planarring or ring system comprising delocalized electrons. These aromaticgroups are preferably monocyclic (e.g., furan or benzene), bicyclic, ortricyclic groups containing from 5 to 14 atoms in the ring portion. Theterm “aromatic” encompasses “aryl” groups defined below.

The terms “aryl” or “Ar” as used herein alone or as part of anothergroup denote optionally substituted homocyclic aromatic groups,preferably monocyclic or bicyclic groups containing from 6 to 10 carbonsin the ring portion, such as phenyl, biphenyl, naphthyl, substitutedphenyl, substituted biphenyl, or substituted naphthyl.

The terms “carbocyclo” or “carbocyclic” as used herein alone or as partof another group denote optionally substituted, aromatic ornon-aromatic, homocyclic ring or ring system in which all of the atomsin the ring are carbon, with preferably 5 or 6 carbon atoms in eachring. Exemplary substituents include one or more of the followinggroups: hydrocarbyl, substituted hydrocarbyl, alkyl, alkoxy, acyl,acyloxy, alkenyl, alkenoxy, aryl, aryloxy, amino, amide, acetal,carbamyl, carbocyclo, cyano, ester, ether, halogen, heterocyclo,hydroxy, keto, ketal, phospho, nitro, and thio.

The terms “halogen” or “halo” as used herein alone or as part of anothergroup refer to chlorine, bromine, fluorine, and iodine.

The term “heteroatom” refers to atoms other than carbon and hydrogen.

The term “heteroaromatic” as used herein alone or as part of anothergroup denotes optionally substituted aromatic groups having at least oneheteroatom in at least one ring, and preferably 5 or 6 atoms in eachring. The heteroaromatic group preferably has 1 or 2 oxygen atoms and/or1 to 4 nitrogen atoms in the ring, and is bonded to the remainder of themolecule through a carbon. Exemplary groups include furyl, benzofuryl,oxazolyl, isoxazolyl, oxadiazolyl, benzoxazolyl, benzoxadiazolyl,pyrrolyl, pyrazolyl, imidazolyl, triazolyl, tetrazolyl, pyridyl,pyrimidyl, pyrazinyl, pyridazinyl, indolyl, isoindolyl, indolizinyl,benzimidazolyl, indazolyl, benzotriazolyl, tetrazolopyridazinyl,carbazolyl, purinyl, quinolinyl, isoquinolinyl, imidazopyridyl, and thelike. Exemplary substituents include one or more of the followinggroups: hydrocarbyl, substituted hydrocarbyl, alkyl, alkoxy, acyl,acyloxy, alkenyl, alkenoxy, aryl, aryloxy, amino, amide, acetal,carbamyl, carbocyclo, cyano, ester, ether, halogen, heterocyclo,hydroxy, keto, ketal, phospho, nitro, and thio.

The terms “heterocyclo” or “heterocyclic” as used herein alone or aspart of another group denote optionally substituted, fully saturated orunsaturated, monocyclic or bicyclic, aromatic or non-aromatic groupshaving at least one heteroatom in at least one ring, and preferably 5 or6 atoms in each ring. The heterocyclo group preferably has 1 or 2 oxygenatoms and/or 1 to 4 nitrogen atoms in the ring, and is bonded to theremainder of the molecule through a carbon or heteroatom. Exemplaryheterocyclo groups include heteroaromatics as described above. Exemplarysubstituents include one or more of the following groups: hydrocarbyl,substituted hydrocarbyl, alkyl, alkoxy, acyl, acyloxy, alkenyl,alkenoxy, aryl, aryloxy, amino, amide, acetal, carbamyl, carbocyclo,cyano, ester, ether, halogen, heterocyclo, hydroxy, keto, ketal,phospho, nitro, and thio.

The terms “hydrocarbon” and “hydrocarbyl” as used herein describeorganic compounds or radicals consisting exclusively of the elementscarbon and hydrogen. These moieties include alkyl, alkenyl, alkynyl, andaryl moieties. These moieties also include alkyl, alkenyl, alkynyl, andaryl moieties substituted with other aliphatic or cyclic hydrocarbongroups, such as alkaryl, alkenaryl and alkynaryl. Unless otherwiseindicated, these moieties preferably comprise 1 to 20 carbon atoms.

The term “protecting group” as used herein denotes a group capable ofprotecting an oxygen atom (and hence, forming a protected hydroxy),wherein the protecting group may be removed, subsequent to the reactionfor which protection is employed, without disturbing the remainder ofthe molecule. Exemplary protecting groups include ethers (e.g., allyl,triphenylmethyl (trityl or Tr), p-methoxybenzyl (PMB), p-methoxyphenyl(PMP)), acetals (e.g., methoxymethyl (MOM), β-methoxyethoxymethyl (MEM),tetrahydropyranyl (THP), ethoxy ethyl (EE), methylthiomethyl (MTM),2-methoxy-2-propyl (MOP), 2-trimethylsilylethoxymethyl (SEM)), esters(e.g., benzoate (Bz), allyl carbonate, 2,2,2-trichloroethyl carbonate(Troc), 2-trimethylsilylethyl carbonate), silyl ethers (e.g.,trimethylsilyl (TMS), triethylsilyl (TES), triisopropylsilyl (TIPS),triphenylsilyl (TPS), t-butyldimethylsilyl (TBDMS), t-butyldiphenylsilyl(TBDPS) and the like. A variety of protecting groups and the synthesisthereof may be found in “Protective Groups in Organic Synthesis” by T.W. Greene and P. G. M. Wuts, John Wiley & Sons, 1999.

The “substituted hydrocarbyl” moieties described herein are hydrocarbylmoieties which are substituted with at least one atom other than carbon,including moieties in which a carbon chain atom is substituted with aheteroatom such as nitrogen, oxygen, silicon, phosphorous, boron, or ahalogen atom, and moieties in which the carbon chain comprisesadditional substituents. These substituents include alkyl, alkoxy, acyl,acyloxy, alkenyl, alkenoxy, aryl, aryloxy, amino, amide, acetal,carbamyl, carbocyclo, cyano, ester, ether, halogen, heterocyclo,hydroxy, keto, ketal, phospho, nitro, and thio.

The term “treating,” as used herein, refers to inhibiting or alleviatingthe symptoms of the disease or disorder; reversing, inhibiting, orslowing the progression of the disease or disorder; and/or preventing ordelaying the onset of the disease or disorder. The term “treatment”, asused herein, unless otherwise indicated, refers to the act of treatingas “treating” is defined immediately above.

When introducing elements of the present invention or the preferredembodiments(s) thereof, the articles “a”, “an”, “the” and “said” areintended to mean that there are one or more of the elements. The terms“comprising”, “including” and “having” are intended to be inclusive andmean that there may be additional elements other than the listedelements.

Having described the invention in detail, it will be apparent thatmodifications and variations are possible without departing from thescope of the invention defined in the appended claims.

EXAMPLES

The following examples illustrate various embodiments of the invention.

Example 1: Toll-Like Receptor Screening

Stimulation of TLRs 2, 3, 4, 5, 7, 8, and 9 was determined by assessingactivation of the transcription factor NF-κB in HEK293 cells that wereengineered to express the corresponding receptors. Assessment of TLRstimulation was based on the use of an NF-κB-inducible secreted alkalinephosphatase (SEAP) reporter system in which the SEAP reporter was underthe control of a promoter inducible by NF-κB. Thus, the degree ofactivation of TLRs can be indirectly quantified spectrophotometricallyby measuring the amount of the SEAP reporter that is produced.

General Procedure.

The appropriate TLR-expressing cells were plated in Growth Medium in a96-well plate (25,000-50,000 cells/well). The cells were stimulated withthe appropriate positive control ligand or no ligand was added (negativecontrol). The positive control ligands were: HKLM was used to stimulateTLR2; poly(I:C) was used to stimulate TLR3; LPS was used to stimulateTLR4; Flagellin was used to stimulate TLR5; CL097 was used to stimulateTLR7; CL075 was used to stimulate TLR8; CpG oligodeoxynucleotide (ODN)2006 was used to stimulate TLR9. To test whether (+)-morphinans couldblock that activation of the TLR, the cells were pretreated with anantagonist for 30 minutes prior to addition of the positive controlligand. For this, 20 μL of the stock test compound solution (100 μM inH₂O) was added to give a total volume of 200 μL. The antagonists testedwere (+)-naloxone, (+)-naltrexone, sinomenine, and dihydrosinomenine;the final concentration of each was 10 μM. After a 16-20 hr incubationperiod at 37° C. in a CO₂ incubator, 20 μL of the cell culturesupernatant was added to 180 μL of QUANTI-Blue™ Media (InvivoGen, SanDiego, Calif.), and the resulting solutions were incubated at 37° C. foran additional 1-3 hours according to the manufacturer's instructions.The OD's of the samples were then read at 650 nm on a Beckman Coulter AD340C Absorbance Detector.

Results.

The results of the antagonist screening experiments are presented inTable 1 and FIG. 1A to FIG. 1G. Each antagonist inhibited the activityof the TLRs. The greatest inhibition, however, was observed with TLR9(see FIG. 1G). These data indicate that TLR9 is the primary target of(+)-morphinans.

TABLE 1 Antagonist Screening Average % Inhibition at Toll-Like ReceptorsCompound TLR2 TLR3 TLR4 TLR5 TLR7 TLR8 TLR9 (+)-Naloxone 24 27 25 17 1824 45 (+)-Naltrexone 21 22 24 22 16 19 51 Sinomenine 30 28 24 19 20 2055 Dihydrosinomenine 27 40 23 16 11 21 42

Example 2: TLR9 Screening of (+)-Morphinan Library

The secreted alkaline phosphatase reporter is under the control of apromoter inducible by the transcription factor NF-κB. A library of morethan 100 (+)-morphinan compounds was screened for TLR9 antagonistactivity by assessing NF-κB activation in the HEK293 cells expressingTLR9. This reporter gene allows the monitoring of signaling through theTLR, based on the activation of NF-κB.

In a 96-well plate (200 μL total volume) containing 50,000 cells/well,20 μL of each sample compound was added to wells, in triplicate,followed by a 30 minute incubation at 37° C. and 5% CO₂. After the 30minutes of incubation, 20 μL of the activator ODN2006 was added to eachwell. The media added to the wells was designed for the detection ofNF-κB induced SEAP expression. After a 16-20 hr incubation period, theOD at 650 nm was read on a Beckman Coulter AD 340C Absorbance Detector.The results of the experiments are shown in Table 2. All compounds weretested at concentrations of 10 μM and 100 nM.

TABLE 2 TLR9 Inhibition OD 650 nm % % (mean ± Inhibition* OD 650 nmInhibition* Compound sd) (mean) (mean ± sd) (mean) ID Chemical Structure10 μM 100 nM II-64

0.98 ± 0.01 31 1.31 ± 0.14 8 II-73

0.96 ± 0.08 32 1.35 ± 0.13 5 I-30

0.94 ± 0.05 33 1.25 ± 0.11 12 I-9

1.03 ± 0.01 28 1.50 ± 0.10 −6 I-25

0.52 ± 0.07 63 1.08 ± 0.05 24 I-18

1.01 ± 0.06 29 1.40 ± 0.14 1 II-26

0.12 ± 0.00 92 1.08 ± 0.06 24 II-24

0.09 ± 0.01 94 0.65 ± 0.04 54 II-25

0.21 ± 0.06 85 1.33 ± 0.07 6 II-17

0.39 ± 0.01 72 1.13 ± 0.10 21 II-28

0.87 ± 0.03 39 1.32 ± 0.08 7 II-29

0.33 ± 0.03 77 1.36 ± 0.17 4 II-31

1.37 ± 0.11 3 1.40 ± 0.04 2 II-57

1.15 ± 0.03 19 0.84 ± 0.02 41 II-59

1.41 ± 0.13 1 1.19 ± 0.07 17 II-60

0.17 ± 0.03 88 1.24 ± 0.04 13 II-83

1.11 ± 0.09 22 1.40 ± 0.13 1 II-84

1.51 ± 0.08 −6 1.44 ± 0.06 −1 II-85

1.45 ± 0.04 −2 1.45 ± 0.04 −2 II-30

1.13 ± 0.06 20 1.34 ± 0.11 5 II-33

1.37 ± 0.03 3 1.35 ± 0.15 5 II-86

0.13 ± 0.03 91 1.15 ± 0.11 19 II-87

0.45 ± 0.01 69 1.28 ± 0.06 10 II-88

0.13 ± 0.01 91 1.12 ± 0.15 21 II-34

0.94 ± 0.08 34 1.39 ± 0.05 2 II-22

0.75 ± 0.06 47 1.31 ± 0.19 8 II-89

1.08 ± 0.02 24 1.53 ± 0.10 −8 I-26

0.19 ± 0.02 87 1.15 ± 0.03 19 II-70

0.79 ± 0.03 45 1.12 ± 0.04 21 II-79

1.06 ± 0.11 25 1.07 ± 0.03 25 III-2

0.42 ± 0.04 71 1.13 ± 0.02 21 III-3

0.13 ± 0.01 91 1.03 ± 0.03 27 III-4

0.11 ± 0.01 92 1.04 ± 0.04 27 III-5

0.29 ± 0.01 80 1.11 ± 0.02 22 II-35

0.34 ± 0.03 76 1.04 ± 0.08 27 II-36

0.14 ± 0.01 90 1.14 ± 0.02 20 II-37

0.18 ± 0.06 87 1.13 ± 0.07 20 II-38

0.12 ± 0.01 92 0.86 ± 0.01 40 I-22

0.10 ± 0.01 93 0.93 ± 0.15 34 II-41

1.02 ± 0.09 28 1.27 ± 0.03 11 II-42

0.14 ± 0.02 90 1.00 ± 0.03 29 II-43

0.09 ± 0.00 94 0.10 ± 0.01 93 II-44

1.03 ± 0.12 27 1.18 ± 0.02 17 II-75

0.91 ± 0.11 36 1.08 ± 0.02 24 II-76

0.10 ± 0.01 93 0.16 ± 0.01 89 II-77

0.98 ± 0.02 37 1.24 ± 0.04 13 II-71

0.10 ± 0.00 93 0.19 ± 0.01 87 II-72

0.49 ± 0.01 65 1.10 ± 0.02 22 II-40

1.37 ± 0.07 4 1.41 ± 0.13 0 II-46

1.30 ± 0.02 8 1.24 ± 0.02 13 II-47

0.78 ± 0.07 45 1.18 ± 0.13 17 II-48

1.23 ± 0.02 13 1.43 ± 0.05 0 II-48

1.06 ± 0.02 25 1.21 ± 0.02 15 II-51

0.84 ± 0.05 41 1.20 ± 0.04 16 II-55

1.17 ± 0.08 18 1.46 ± 0.08 −3 II-74

0.12 ± 0.00 92 0.61 ± 0.03 57 II-90

0.17 ± 0.05 88 1.39 ± 0.03 2 II-78

0.11 ± 0.01 92 0.14 ± 0.02 90 II-82

0.49 ± 0.03 65 1.37 ± 0.02 4 II-62

0.18 ± 0.01 87 1.34 ± 0.05 6 II-63

0.13 ± 0.01 91 0.23 ± 0.07 84 I-2

0.14 ± 0.02 90 1.10 ± 0.06 22 I-1

0.21 ± 0.01 85 1.18 ± 0.10 17 I-3

0.11 ± 0.00 92 1.12 ± 0.01 21 I-7

0.16 ± 0.01 89 1.14 ± 0.03 20 II-1

0.19 ± −0.2 86 0.97 ± 0.04 32 II-2

0.12 ± 0.01 92 1.23 ± 0.05 13 II-4

0.11 ± 0.01 92 1.18 ± 0.06 17 I-8

0.11 ± 0.01 92 1.13 ± 0.07 20 I-11

1.98 ± 0.09 −39 1.49 ± 0.04 −5 I-12

0.13 ± 0.01 91 1.19 ± 0.07 16 I-13

0.13 ± 0.01 91 1.34 ± 0.12 6 I-27

0.10 ± 0.00 93 1.13 ± 0.04 20 II-15

0.23 ± 0.03 84 1.22 ± 0.06 14 II-16

1.24 ± 0.11 13 1.43 ± 0.01 −1 II-23

0.53 ± 0.05 63 1.16 ± 0.01 18 II-21

0.23 ± 0.01 84 1.40 ± 0.02 2 I-5

0.87 ± 0.04 38 1.31 ± 0.09 8 I-4

0.46 ± 0.05 68 1.17 ± 0.06 18 I-6

0.12 ± 0.02 91 1.29 ± 0.07 9 I-5

0.14 ± 0.00 90 1.11 ± 0.03 22 II-11

0.44 ± 0.05 69 1.22 ± 0.10 14 II-14

0.13 ± 0.00 91 1.18 ± 0.05 17 II-7

0.13 ± 0.02 91 1.32 ± 0.08 7 II-91

0.09 ± 0.01 93 0.12 ± 0.01 92 II-92

0.73 ± 0.06 48 1.09 ± 0.03 23 II-93

0.11 ± 0.01 92 0.12 ± 0.00 91 II-21

0.09 ± 0.01 93 0.19 ± 0.02 87 II-94a

1.16 ± 0.08 18 1.13 ± 0.04 20 II-94b

1.30 ± 0.05 8 1.17 ± 0.03 17 II-9

1.00 ± 0.02 29 1.25 ± 0.09 12 II-20

1.07 ± 0.04 25 1.29 ± 0.02 9 II-19

0.15 ± 0.01 90 1.19 ± 0.03 16 II-8

1.33 ± 0.06 7 1.15 ± 0.07 19 II-53a

1.32 ± 0.13 7 1.22 ± 0.05 14 II-53b

1.00 ± 0.03 29 1.41 ± 0.09 1 II-74

1.26 ± 0.06 11 1.19 ± 0.06 16 II-95

0.37 ± 0.03 74 1.11 ± 0.04 22 II-96

0.42 ± 0.02 71 1.15 ± 0.06 19 II-97

1.42 ± 0.07 0 1.22 ± 0.02 14 II-98

0.56 ± 0.03 60 1.02 ± 0.02 28 II-99

0.14 ± 0.01 90 1.32 ± 0.00 7 II-100

1.45 ± 0.11 −2 1.16 ± 0.02 18 II-12

0.81 ± 0.03 43 1.12 ± 0.08 21 II-101

0.94 ± 0.02 34 1.35 ± 0.06 5 III-1

0.67 ± 0.01 53 1.34 ± 0.06 6 II-39

1.06 ± 0.06 25 1.23 ± 0.03 14 II-10

0.10 ± 0.01 93 0.24 ± 0.02 83 I-16

0.91 ± 0.04 36 1.50 ± 0.01 −5 I-28

0.12 ± 0.01 91 1.24 ± 0.02 13 I-10

1.04 ± 0.08 27 1.32 ± 0.04 7 I-29

0.11 ± 0.00 92 1.06 ± 0.03 25 II-61

1.22 ± 0.07 14 1.23 ± 0.02 13 *Control (no activator) = 0.14 OD 650 nm(n = 12); Activator (ODN2006) = 1.42 OD 650 nm (n = 12)

This screening experiment identified those (+)-morphinan compoundscomprising the greatest TLR9 antagonist activity. Even at aconcentration as low as 100 nM, some compounds inhibited TLR9 in excessof 90%.

Example 3: Analgesic Assessment of (+)-Naloxone on Mechanical Allodynia

Neuropathic pain affects approximately 1% of the U.S. population and isextremely difficult to manage. Usually the pain is chronic, severe, andfails to respond to traditional analgesic drugs. Fortunately, one of themost widely used animal models for neuropathic pain closely mimics thepain endured by patients. In this model, termed the chronic constrictioninjury (CCI) or Bennett model, four closely spaced ligatures tiedloosely around the sciatic nerve of a rat cause demyelination of thenerve, resulting in spontaneous pain, such as prolonged paw elevationand licking of the ligated paw. Induced pain in the nerve-injured limbcan be measured using Von Frey filaments, applied to the plantar surfaceof the paw to test for responses to non-noxious tactile stimulation. Theonset of this heightened mechanical allodynia is quite rapid and itpersists for 2-3 months.

In this example, the CCI ligation was performed on the left sciaticnerve in three groups of rats. Von Frey testing of mechanical allodyniawas performed on both paws on Day 14 to determine the analgesic efficacyof the test agent. The testing was performed pre-dosing, and at 30 and90 minutes post-dosing.

Animals.

A total of thirty-four (34) male Sprague-Dawley rats were ordered fromHarlan Sprague-Dawley. The animals were specific pathogen free andweighed approximately 175-200 grams upon arrival. A visual healthinspection was performed on each animal to include evaluation of thecoat, extremities and abnormal signs in posture or movement. Animalswere individually identified with a unique ear tag assigned at receipt.The animals were individually housed in clear polycarbonate plasticcages and received enrichment in the way of Enrich-o-cobs bedding. Cagecards were affixed to their cages that identified study number, animalnumber, treatment designation, species/strain, and gender. The animalswere acclimated for 5 days prior to the commencement of the experimentalprocedures. The room number in which the animals were housed throughoutthe study period was detailed in the study records. The temperature wasmaintained at 18-26° C. (64-79° F.) with a relative humidity of 30-70%.Temperature and humidity were monitored and daily minimums and maximumsrecorded.

Treatment Groups.

The animals were allocated to treatment groups based on their baselineVon Frey data, measured prior to surgery. The mechanical allodyniascores for each group were reviewed to ensure that the mean values andstandard deviation satisfied the assumption of homogeneity. Table 3presents the treatment groups. Thirty-one (31) animals were used on thestudy. Thirty (30) animals were initially allocated to treatment groups,and the remaining four (4) animals were held as spares. One spare wasthen used to replace an animal that died during surgery. Body weightswere taken one day after arrival, prior to surgery and weeklythereafter. On Day 14, after the final behavioral testing, the animalswere euthanized by carbon dioxide asphyxiation. No necropsy wasperformed nor tissues collected.

TABLE 3 Treatment Groups Descrip- Test Route/ Dose #/ Group tion ArticleFrequency mg/kg Group 1 Bennett Vehicle S.C. 0 10 surgery injection 2Bennett Gabapentin I.P.  100 mg/kg 10 surgery injection 3 Bennett (+)-S.C. 66.7 mg/kg 10 surgery Naloxone•HCl injection

Surgery.

All surgeries were performed under aseptic conditions. Prior to surgery,the rats were sedated using inhaled Isoflurane anesthetic. The left legwas shaved and prepped. The common sciatic nerve was exposed and freedfrom adherent tissue at mid-thigh by separating the muscle (bicepsfemoris) by blunt dissection. Proximal to the sciatic nerve'strifurcation; approximately 7 mm of nerve was freed from the adheringtissue. Four ligatures, approximately 1 mm apart, were tied looselyaround the nerve using 6.0 chromic catgut. Each of the sutures was tiedloosely with a square knot around the sciatic nerve. A brief twitch inthe muscle surrounding the exposure was an indicator of the desireddegree of constriction. The site was then closed using the appropriatesuture material. Post-operative care and observations were carried outuntil the animal recovered consciousness. Animals were observed fordragging (from inadvertent surgical damage) on Days 1 & 3post-operatively and daily for signs of ill health and general wellbeing.

Behavioral Testing.

The animals underwent a series of behavioral testing for neuropathicpain assessment. A Von Frey test of mechanical allodynia was performedon both hind paws prior to surgery to achieve a baseline measurement forrandomization and then following surgery on Day 14 (test articleefficacy) according to Table 4.

TABLE 4 Behavioral Testing Schedule Day Time point 0 Baseline (forrandomization) 14 Pre-dose 14 30 minutes post-dose 14 90 minutespost-dose

Mechanical Allodynia.

Twice prior to surgery, the animals were acclimated to the allodyniaapparatus. This habituated the rats to the testing devices tofamiliarize them with the apparatus so they were calm at the time oftesting. The test for mechanical allodynia was used to assess theanti-nociceptive properties of analgesic compounds. Animals were firsthabituated to the testing chamber so that they were calm enough fortheir pain threshold to be assessed. A technician blind to the treatmentgroups applied light pressure to both hindpaws of the rat using a seriesof graded nylon filaments (Von Frey filaments) of increasing diameter.The filaments were pressed perpendicularly against the ventral surfaceof the paw until they bent and the rat responded by withdrawing its pawwhen this was considered painful. Threshold allodynia was determinedusing the Chaplan up-down method which provided the precise force forwithdrawal for each rat using a psychophysical scale of testing. Boththe left and right hindpaws were tested at each time point. The order oftesting was the ipsilateral (affected) limb followed by thecontralateral limb. There were approximately 20 minutes in betweentesting the two limbs.

Dosing.

On Day 14, animals in Groups 1 and 3 received an S.C. injection ofeither vehicle or (+)-naloxone.HCl according to Table 3. They were dosedat 1 mL/kg. On Day 14, animals in Group 2 received an I.P. injection ofGabapentin according to Table 3. They were also dosed at 1 mL/kg.

Statistics.

The allodynia data were compared among groups and between paws and timepoints with a 3×2 ANOVA. When data were significant (p<0.05), aBonferroni post-hoc test was applied to determine individual groupdifferences.

Results.

The mechanical allodynia testing was performed prior to surgery(baseline), and at Day 14 post-surgery at pre-dosing, and 30 and 90 minpost-dosing. The baseline data are not included in the figures, but allanimals achieved the maximal score of 17 g, indicating no sensitivity tothe highest force of filament used in this test. At pre-dose testing onDay 14, there were some animals that continued to respond with a scoreof 17 g. This indicated that the surgery had not been effective atcausing neuropathic pain in this subset of animals, and therefore, theywere removed from the data set, and excluded from statistical analyses.The final sample sizes and the allodynia data are presented in FIG. 2Afor the left paw (affected) and FIG. 2B (unaffected) for the right paw.

The data in FIG. 2A demonstrate a significant allodynia in the left pawfor all three groups in relation to the pre-surgery baseline data(p<0.01 for treatment, p<0.001 for time). After dosing, both Gabapentinand (+)-naloxone were effective at significantly reducing the allodyniaat both 30 and 90 min. The right paw data are depicted in FIG. 2B andthese data reveal that there was no allodynia in the right paw asexpected in this model. There were no significant group differences ordifferences across time with the exception of the Gabapentin group,which demonstrated a higher threshold for mechanical allodynia testingat the 90 min timepoint than was observed in the vehicle group. This isnot surprising, given that Gabapentin was administered peripherally andhas a robust analgesic effect bilaterally.

Conclusions.

This study examined the efficacy of the test agent, (+)-naloxone, toreduce allodynia in the left paw of animals that achieved neuropathicpain when tested on Day 14 post-surgery. The results showed thatsignificant analgesia was achieved by (+)-naloxone at both timepointstested, which was similar to the results of the positive control,Gabapentin. Thus, (+)-naloxone.HCl was an effective analgesic forreversing neuropathic pain over a timecourse of 30 and 90 minpost-dosing on Day 14 using a dose of 66.7 mg/kg.

Example 4: Evaluation of (+)-Morphinans in Animal Model of NeuropathicPain

The following example was designed to evaluate the effectiveness ofseveral specific (+)-morphinans to reverse allodynia, as assessed by vonFrey testing, in rats subjected to chronic constriction injury (CCI).

Allodynia was induced in a group of (male, Sprague-Dawley) ratsessentially as described above in Example 3. Prior to surgery, animalswere tested to establish their baseline von Frey scores. Any animal witha Log absolute von Frey score lower than 4.65 on either leg was excludedfrom the study. CCI surgery was performed on the left sciatic nerve asdetailed above. On days 4 and 14 post-surgery, the animals were testedusing the von Frey test of mechanical allodynia to ensure that allodyniawas established.

Beginning at about day 14 post-surgery, the animals were dosed once aday for 4 days. Compounds II-25, II-93, II-62, and III-2 were tested.Table 5 presents the treatment groups. All drugs were administeredintraperitoneally (i.p.). Allodynia was tested prior to and again 3 hrsafter each dosing. Drug administration and testing was done blindly.After the 4^(th) day of dosing, the animals were sacrificed, cardiacperfusion was performed, and spinal cords were removed and stored forfurther analysis.

TABLE 5 Treatment groups Condition Dose #/group Control - saline 0 46Gabapentin 100 mg/kg  24 (+)-naloxone - ultra low 0.071 mg/kg  16(+)-naloxone -low 0.71 mg/kg  8 (+)-naloxone - medium 7.1 mg/kg 8(+)-naloxone - high  71 mg/kg 8 Cmpd II-25 - ultra low 0.073 mg/kg  8Cmpd II-25 - low 0.73 mg/kg  8 Cmpd II-25 - medium 7.3 mg/kg 9 CmpdII-25 - high  73 mg/kg 10 Cmpd II-93 - ultra low 0.079 mg/kg  8 CmpdII-93 - low 0.79 mg/kg  8 Cmpd II-93 - medium 7.9 mg/kg 8 Cmpd II-93 -high  79 mg/kg 8 Cmpd II-62 - ultra low 0.1 mg/kg 8 Cmpd II-62 - low 1.0mg/kg 8 Cmpd II-62 - medium 10.1 mg/kg  8 Cmpd II-62 - high 101 mg/kg  7Cmpd III-2 - ultra low 0.073 mg/kg  8 Cmpd III-2 - low 0.73 mg/kg  8Cmpd III-2 - medium 7.3 mg/kg 8 Cmpd III-2 - high  73 mg/kg 8

All data were analyzed using 2-way ANOVA's with Bonferroni's post hoctest compared to saline unless otherwise stated. Area under curve (AUC)was calculated using Graphpad Prism and comparisons were made using1-way ANOVA with Dunnet's post hoc test.

Results.

All groups of rats developed allodynia. FIGS. 3A and 3B presents arepresentative plot of the development of allodynia in which the vonFrey scores are plotted at day 0 (prior to surgery) and days 4 and 14post-surgery in various treatment groups. None of the rats treated with(+)-naloxone displayed any reversal of allodynia at any dose (data notshown). It appears, therefore, that (+)-naloxone is not effective atreversing allodynia 3 hours post dose using the current protocol.

Compound II-25 showed a significant overall effect on the reversal ofallodynia bilaterally compared to saline after the medium (P<0.05) orthe high (P<0.05) dose. FIG. 4 presents time courses of the allodyniaresponses (i.e., pre-dose and post-dose) in the ipsilateral (panels A-D)and contralateral paws (panels E-H) for each of the four days of dosingwith the different doses of Compound II-25. The effect was mostprominent on Day 1 of dosing in rats treated with the high dose (3 hrsP<0.05) and Day 3 of dosing in rats treated with the medium dose (51 hrsP<0.05). The under the curve analysis is presented in FIGS. 5A and 5B.From this plot, it is apparent that doses of 7.3 mg/kg and 73 mg/kgsignificantly reversed allodynia. It was observed that 5-6 of theanimals (later revealed as having been treated with the high dose ofCompound II-25) appeared sleepier than the other rats on at least onepost dosing occasion.

Compound II-93 significantly reversed allodynia in the ipsilateral pawafter the high dose only (P<0.01). The effect was most prominent on Days2 and 3 of dosing (27 hrs P<0.01; 51 hrs P<0.01) (see FIG. 6, panelsA-D). The under the curve analysis is plotted in FIG. 7A and FIG. 7B, inwhich the significant reversal of allodynia in the ipsilateral paw (seeFIG. 7A) was detected after treatment with the high dose (i.e., 79mg/kg) of Compound II-93. Throughout the testing, many of the rats wereobserved to be sleepy or sedated, and some rats had some swelling of theipsilateral paw. After unblinding, it was revealed that 7 of the animalstreated with the high dose of compound II-93 displayed sedation on atleast one post dosing occasion and had swelling of the ipsilateral paw;and that two rats treated with the medium dose and one rat treated withthe low dose showed some sedation.

Compound II-62 did not significantly reverse allodynia at any dose (datanot shown). The area under the curve analysis demonstrated that therewas no overall effect of any dose of compound II-62 compared to saline,but time course analysis revealed that there was a slight reversal ofallodynia on the ipsilateral paw on Day 4 of dosing, however. Afterunblinding, it was revealed that one rat treated with the low dose, onetreated with the medium dose, and two rats treated with the high dose ofcompound II-62 appeared sleepier than the other rats on at least onepost dosing occasion. It was also observed that injection of the highdose of compound II-62 appeared to irritate the rats when it wasinjected. Some of the rats administered this dose showed swelling aroundthe site of injection the next day.

Compound III-2 had no effect on allodynia at any dose (data not shown).There were no significant effects of this compound compare to saline inany of the statistical tests conducted. Two rats later revealed to havebeen treated with the low dose of compound III-2 had increased levels ofsleepiness during one of the post dosing testing sessions.

In summary, this study revealed that two of the tested (+)-morphinans,i.e., compounds II-25 and II-93, were effective at reversing mechanicalallodynia.

Example 5: Evaluation of (+)-Morphinans to Inhibit/ReduceAcetaminophen-Induced Hepatotoxicity

Acetaminophen-induced liver damage is the most common cause of death dueto acute liver failure. Treatment with a (+)-morphinan may reduce orprevent acute liver injury induced by acetaminophen (APAP). Theeffectiveness of some of the (+)-morphinan compounds disclosed herein toreduce liver injury after exposure to a toxic dose of APAP was tested inmouse model of APAP-induced toxicity.

For this, APAP-induced liver toxicity was induced in male C57BL/6 miceaged between 8-10 weeks by a single intraperitoneal (ip) injection ofAPAP in PBS at a dose of 500 mg/Kg (Imaeda et al. (2009) J. Clin.Invest. 119(2):305-314). The ability of (+)-morphinans to reduce livertoxicity was tested in the following groups:

1) No APAP and PBS

2) APAP and PBS

3) No APAP and (+)-morphinan

4) APAP and (+)-morphinan

The (+)-morphinan (or PBS) was either administered as a singlesubcutaneous (sc) or ip injection concurrent with the APAP injection orone hour prior to the APAP injection. Table 6 presents the(+)-morphinans tested.

TABLE 6 Test compounds and doses. Test Agent Dose Timing (+)-naloxone 60mg/kg, Concurrent ip (PBS) Cmpd II-78 30 mg/kg, One hr prior sc (PBS)Cmpd II-24 60 mg/kg, One hr prior sc (PBS) Cmpd II-93 60 mg/kg, One hrprior sc (PBS) Cmpd III-2 30 mg/kg, One hr prior sc (50% EtOH/NS)

The mice were sacrificed 12 hours post-APAP treatment. Liver sampleswere stained with H&E and analyzed for signs of necrosis and hemorrhage.Histology scores were assigned using the standard rating scale. Thelevels of serum alanine transaminase (ALT) were also measured in eachanimal using a standard assay.

Animals administered APAP alone had significant liver injury asevidenced by increased histology scores and elevated ALT levels.(+)-Naloxone administered at the same time as the toxic dose of APAPprovided essentially no protection. As shown in FIG. 8 (panels A, B),(+)-naloxone reduced the ALT levels, but did not improve the histologyscores for necrosis and hemorrhage.

Compound II-78 provided protection from APAP-induced toxicity. CompoundII-78 protected the liver from necrosis and hemorrhage (P<0.05)—thehistology scores were reduced to about half of those treated only withAPAP (see FIG. 9, panel A). Similarly, ALT levels were reducedsignificantly (P<0.05) by pretreatment with compound II-78 (see FIG. 9,panel B).

None of compounds II-24, II-93, or III-2 provided any protection fromAPAP-induced toxicity (data not shown). For each compound, the histologyscores were about the same as those only exposed to APAP, and ALT levelswere similar between the two groups.

Example 6: Evaluation of (+)-Morphinans to Inhibit/Reduce InflammationUsing EAE Model

The effectiveness of (+)-morphinans to reduce inflammation was testedusing the EAE (Experimental Autoimmune Encephalomyelitis (EAE) Lewis ratmodel. For this, six groups of 10 rats/group of female Lewis rats (7weeks old; ˜200 g) were used in the study. The groups included a vehicle(20% 2-hydroxypropyl, β-cyclodextrin, HBC), a positive control(prednisolone), and four test (+)-morphinans. The groups are presentedbelow:

1. Vehicle: 2.5 ml/kg of 20% HBC; 80% PBS 2. Prednisolone: 4.5 mg/kg(20% HBC; 80% PBS), ip 3. Cmpd II-24 20 mg/kg (sterile, dd H₂O), sc 4.Cmpd II-38 20 mg/kg (sterile, dd H₂O), sc 5. Cmpd II-78 20 mg/kg(sterile, dd H₂O), sc 6. Cmpd II-98  5 mg/kg (sterile, dd H₂O), sc

Each rat was injected with 0.05 ml of an emulsion of Mycobacteriumtuberculosis H37 and guinea pig spinal cord into each main foot pad ofthe rear paws using conscious restraint. The compounds (or vehicle) wereadministered twice daily (bid). The first dose was give prior to EAEinduction. Dosing continued for 20 days.

Body weight, paw thickness and clinical signs of EAE were measured foreach rat 2 times per week starting the 2nd week of treatment andcontinued until the end of study. Clinical scoring was conductedaccording to the table below. Rats in which scores were not clear cutwere give half scores, such as 3.5. Statistics were conducted on theclinical scores for the period in which onset of disease was observeduntil recovery. A Repeated Measures One Way ANOVA followed by aNewman-Keuls posthoc test if significant was conducted (Graphpad Prism5, La Jolla, Calif.). Data presented as Mean±SEM.

EAE Score Symptoms 0 Normal 1 Limp tail 2 Incomplete paralysis of one orboth hind limbs (weakness, limping, shaking) 3 Complete paralysis of onehind limb or both hind limbs can move but do not help in movement of thebody 4 Complete paralysis of both hind limbs (back half or rat isdragged around the cage) 5 Complete paralysis of hind limbs and weaknessof one or both forelimbs or moribund, or death

The EAE clinical scores for each group of rats are presented in FIG. 10.Prednisolone (positive control) and Cmpd II-98 (5 mg/kg bidsubcutaneous) significantly reduced EAE clinical scores compared tovehicle control (p<0.05). Initially, Cmpd II-98 was dosed at 20 mg/kgbid, however, acute toxicity was observed. Starting on the second day ofdosing, Cmpd II-98 was delivered at 5 mg/kg bid subcutaneous.

Compd II-24, Cmpd II-38, and Cmpd II-78 were delivered subcutaneous at20 mg/kg bid and did not improve EAE compared to vehicle control.However, Cmpd II-38 did not dissolve into a solution and was deliveredthroughout the study as a suspension. It is not known whether thesuspension dissolved adequately in the body to provide sufficient plasmaconcentration for efficacy.

In summary, Cmpd II-98 had anti-inflammatory effects in the EAE Lewisrat model.

Example 7: Evaluation of (+)-Morphinans to Inhibit/Reduce Tumor Growthand Metastasis

To determine whether (+)-morphinans inhibit pancreatic tumor growth andmetastasis, the following example may be performed. Human pancreatictumor cell line AsPC-1 cells (˜1×10⁶) may be injected into the pancreasof nude mice on Day 0. On Day 10 (when the tumors are about 3-4 mm indiameter), the mice may be divided into at least two treatment groups(˜10 mice per group): (1) control—no treatment; and (2) (+)-morphinantreatment. The control group may be administered PBS or vehicle in amanner similar to the (+)-morphinan treatment group(s). The(+)-morphinan may be administered ip daily for 10-15 days, sc daily for10-15 days, or iv every 4 days for a total of 4 times. On day 24/25, themice may be sacrificed. To determine whether treatment with the(+)-morphinan reduced tumor growth, the tumors may be weighed and/or thevolume of the tumors may be quantified. The occurrence of metastaticloci may be determined in the lungs, liver, and lymph to determine theeffect of the (+)-morphinan on metastasis.

A similar analysis may be performed using a (+)-morphinan in combinationwith another type of chemotherapeutic agent.

Example 8: Evaluation of (+)-Morphinans in Combination withChemotherapeutic Agent to Treat Xenograft Tumor

To determine whether (+)-morphinans increase the efficacy ofchemotherapeutic agents, the following trial may be performed. Femalenude mice (Hsd:Athymic Nude-Foxn1 nu/nu; 5-6 weeks old) may be injectedsc into the right shoulder region with EGFR expressing human tumorcells. The mice may be divided into the following treatment groups (˜10rats/group):

1) Vehicle: saline

2) Cisplatin 6 mg/kg, iv, 3× daily

3) Cisplatin 6 mg/kg iv and (+)-morphinan 20 mg/kg, sc, 3× daily

4) EGFR inhibitor, iv, 3× daily

5) EGFR inhibitor iv and (+)-morphinan 20 mg/kg, sc, 3× daily

6) (+)-morphinan 20 mg/kg, sc, 3× daily

Palpation for tumors may begin 7 days post implantation. Tumors may beobserved and measured 3 times a week. Caliper measures—in mm width(small measure)×length (large measure). Body weight may be recorded onday 1 of the trial (day of cell implantation) and once weekly untilnecropsy. NIH euthanasia guidelines for rodent tumors will be followed(e.g., if tumor diameter exceeds 20 mm, if tumor is ulcerated tumor, iftumor severely restricts the animal's ability to eat, drink, eliminatewastes, breathe, or ambulate, or if animal is becoming emaciated and/orloses more than 20% of pre-study weight).

Example 9: Evaluation of (+)-Morphinan to Reduce Liver TransplantationInduced Reperfusion Injury

Organ transplantation is the only therapeutic option once an organ hasreached the point of end-stage disease, and is the only hope of survivalafter end-stage liver disease. One of the main limitations in organtransplantation is the injury to the donor organ at the time oftransplantation. The bulk of this injury occurs at the time that thetransplant is perfused in the recipient (i.e., ischemia reperfusion (IR)injury). IR injury is an inflammation-mediated injury, and studies inknockout mice have shown that it is greatly reduced in the absence ofTLR9.

The following example tested the utility of a (+)-morphinan (i.e.,compound II-78) to protect against IR injury. Finding that(+)-morphinans reduced or prevented IR injury would increase the changeof organ survival and possibly expand the number of donor organs thatcould be used.

Anesthetized mice were subjected to 1 hour of hepatic ischemia byoccluding the vasculature supplying the left and median lobes (ischemiclobes) of the liver by a microvascular clamp. Thereafter, the bloodsupply was resumed (reperfusion). Some animals were sacrificed after 3hour of reperfusion, and others after 12 hours or reperfusion. Theexperimental group received Cmpd II-78 at a dose of 80 mg/kg givenintraperitoneally 1 hour prior to starting ischemia. The compound wasadministered before the start of ischemia to mimic the presumed clinicaltiming of delivery.

Cellular Response.

Livers were sectioned, stained, examined, and scored (Suzuki scoringsystem) based on the presence and/or severity of sinusoidal congestion,cytoplasmic vacuolization, and necrosis of parenchymal cells, asindicated in the table below. The levels of serum alanine transaminase(ALT) were also measured using a standard assay.

Histological examination of livers showed that after 1 hour of ischemiaand then reperfusion for 3 hours, there was some liver injury (FIG. 11,panel A). There was much more injury, however, after 12 hours ofreperfusion (FIG. 11, panel B). Injury at both time points was reducedby treatment with the (+)-morphinan (FIG. 11, panels A and B). Withliver injury there is a release of cytoplasmic proteins into thecirculation. Serum alanine transaminase (ALT) is a commonly used markerfor liver injury. As shown in FIG. 12, IR injury elevated the levels ofALT, but treatment with the (+)-morphinan reduced liver injury,resulting in lower levels of ALT. Although the (+)-morphinansignificantly reduced liver injury (serum ALT) at the 12 hour time point(FIG. 12, panels B and C), there was a trend towards reduction at the 3hour time point (FIG. 12, panel A).

Molecular Response.

IL-1β, MCP-1, and NLPR3 have pro-inflammatory and tissue damagingproperties, and IL-1R antagonist (IL-1Ra) has anti-inflammatory andpro-reparative functions in the liver. MAP kinases are part of animportant cytoplasmic signaling cascade that are known to be activated(i.e., phosphorylated) by IR. The levels of IL-1β, MCP-1, NLPR3, andIL-1Ra mRNA were determined in whole liver (at the 12 hour time point)using standard techniques (e.g., RT-PCR). The activation of MAP kinaseswas determined in whole liver lysates by Western blotting usingphospho-specific antibodies (12 hour time point).

The levels of IL-1β, MCP-1, and NLPR3 mRNA were increased by IR, but the(+)-morphinan reduced the levels of the pro-inflammatory cytokines (FIG.13, panels A-C). Treatment with the (+)-morphinan also increased thelevels of the anti-inflammatory molecule, IL1Ra (FIG. 13, panel D). FIG.14 (panel A) shows that the (+)-morphinan also reduced the IR-inducedactivation of MAP kinases. The IR-induced activation and cleavage ofTLR9 were also reduced by the (+)-morphinan (FIG. 14, panel B).

TLR9 signalling involves activation of NF-κB and IRF downstreampathways. The (+)-morphinan inhibited both of these pathways followingactivation of TLR9 (but not activation of TLR4 or other receptors). Thetype C TLR9 agonist, ODN2395, and the TL4 agonist, LPS, both induceup-regulation of IL-1β in peritoneal macrophages via activation ofNF-κB. The TLR9, but not the TLR4, response was inhibited by the(+)-morphinan (FIG. 15, panel A). The TLR9 agonist, ODN2395, activatesthe IRF dependent up-regulation of IFN-β in plasmacytoid dendriticcells, and this activation was inhibited by the (+)-morphinan (FIG. 15,panel B). In contrast, poly (dA:dT) activates the cytoplasmic DNAreceptor AIM2, which results in up-regulation of IF-β in plasmacytoiddendritic cells, but this activation was not blocked by the(+)-morphinan (FIG. 15, panel B).

These data show that (+)-morphinans reduce Inflammation-mediated injuryin livers stressed by ischemia-reperfusion, and may be useful duringhepatic resections and liver transplantations.

Example 10: Evaluation of (+)-Morphinan to Reduce IR Injury in High FatDiet (HFD) Model of Ischemia Reperfusion

Significantly more people need liver transplants than there areavailable donor organs. In addition to the scarcity of liver donations,many of the donor livers have a significant degree of steatosis which isknown to increase IR injury. Thus, many otherwise healthy, butsteatotic, livers are rejected for transplantation. If a drug can beshown to reduce IR injury in steatotic livers it would allow use ofsteatotic livers for transplantation and increase the total number ofliver transplants that could be performed. To test this, mice wereplaced on a high fat diet (HFD) for 12 weeks to induce hepaticsteatosis, which was followed by IR injury. For this, mice weresubjected to hepatic ischemia by occluding the vasculature supplying theleft and median lobes (ischemic lobes) of the liver by a microvascularclamp. After 1 hour of ischemia, the blood supply was resumed(reperfusion). Some animals were sacrificed after 3 hour of reperfusion,and others after 12 hours or reperfusion. The experimental groupreceived Cmpd II-78 at a dose of 80 mg/kg given intraperitoneally 1 hourprior to starting ischemia. Cellular and molecular responses wereassessed essentially as described above in Example 9.

Examination of hematoxylin and eosin stained liver sections revealedthat livers from mice on a HFD were more steatotic than those from micefed a regular diet. Suzuki scores are presented in FIG. 16, which showthat compared to mice fed a regular diet there is greater injury at the3 h time point (compare FIG. 16, panel A with FIG. 11, panel A). The(+)-morphinan protected steatotic livers from IR injury at 3 h and 12 hafter reperfusion (FIG. 16, panels A-B), and reduced serum ALT levels(FIG. 17, panels A-C).

The (+)-morphinan also reduced the pro-inflammatory cytokines, IL-1β,MCP-1, and NLPR3 (FIG. 18, panel A-C) and increased the levels of theanti-inflammatory molecule, IL1Ra (FIG. 18, panel D) in steatoticlivers. The IR-induced activation of MAP kinases was inhibited by the(+)-morphinan (FIG. 19, panel A) in steatotic livers. Similarly, theIR-induced activation and cleavage of TLR9 were also reduced by the(+)-morphinan (FIG. 19, panel B) in steatotic livers. These data showthat (+)-morphinans can be used reduce IR injury in steatotic livers.

Example 11: Evaluation of (+)-Morphinan to Reduce Inflammation in AcutePancreatitis Model

Acute pancreatitis can be induced in rodents by administration ofcaerulein, a CCK analog derived from derived from the Australian treefrog Litoria caerulea. To determine whether a (+)-morphinan could reducethe inflammation and/or consequences of pancreatitis, the followingexample was performed. Mice were administered subcutaneously aneffective dose of caerulein and either saline or Cmpd II-78 (30 mg/kg).After the standard period of time, the mice were sacrificed. Pancreatictissue sections were stained and examined for neutrophil infiltration,and pancreatic trypsin, serum amylase, and pancreatic caspase −1activities were measure using standard assays.

The results are shown in FIG. 20. The (+)-morphinan dramatically reducedthe extent of neutrophil infiltration into the pancreatic tissue (seepanel A). Significant reductions in trypsin activation, serum amylase,and caspase-1 activity were observed (see panels B-D). These data showthat (+)-morphinans can be used to treat acute pancreatitis.

What is claimed is:
 1. A method for therapeutically treating a hepaticdisorder mediated by Toll-like receptor 9 (TLR9) in a subject in needthereof, the method comprising administering to the subject a compoundof Formula (IIb) or a pharmaceutically acceptable salt thereof:

wherein: R is hydrogen, alkyl, allyl, cyclopropylmethyl, orcyclobutylmethyl; R^(a) is chosen from hydrogen, alkyl, or phenyl; R^(b)is hydrogen, alkyl, alkylalcohol, alkylcarboxyl,alkylcarboxylalkylester, acyl, acylalkyl, acylaryl, acyloxy,acyloxyalkyl, or acyloxyaryl; and Y is chosen from hydrogen and hydroxy;provided that R is other than allyl when R^(a) is methyl, R^(b) ispropyl; and Y is hydrogen.
 2. The method of claim 1, wherein R iscyclopropylmethyl, R^(a) is hydrogen, R^(b) is methyl, and Y is hydroxy.3. The method of claim 1, wherein R is methyl, R^(a) is methyl, R^(b) ismethylacetylmethylester; and Y is hydrogen.
 4. The method of claim 1,wherein R is allyl, R^(a) is methyl, R^(b) is ethylalcohol; and Y ishydrogen.
 5. The method of claim 1, wherein the hepatic disorder is aninflammatory disorder.
 6. The method of claim 1, wherein the hepaticdisorder is ischemic-reperfusion injury.
 7. The method of claim 1,wherein the hepatic disorder is chosen from hepatic steatosis, hepaticsteatosis post-liver transplantation, hepatitis, autoimmune hepatitis,alcoholic hepatitis, viral hepatitis, non-alcoholic fatty liver disease,ischemic hepatitis, metabolic disorder hepatitis, chronic liverinflammation, hepatic fibrosis, cholestasis, steatosis, hepaticgranulomas, acute or chronic liver transplant rejection and metabolicconditions, hepatocellular carcinoma, cholangiocarcinoma, primarysclerosing cholangitis, cirrhosis, primary biliary cirrhosis, zonalnecrosis, hemochromatosis, Wilson's disease, alpha 1-antitrypsindeficiency, glycogen storage disease type II, Gilbert's syndrome, portalhypertension, portal vein thrombosis, ascites, variceal bleeding, buddchiari, hepatic viral infections, hepatic bacterial infections, hepaticparasitic infections, liver abscess, or a combination thereof.
 8. Amethod for inhibiting hepatic ischemic-reperfusion injury in a subjectin need thereof, the method comprising administering to the subject acompound of Formula (IIb) or a pharmaceutically acceptable salt thereof:

wherein: R is hydrogen, alkyl, allyl, cyclopropylmethyl, orcyclobutylmethyl; R^(a) is hydrogen, alkyl, or phenyl; R^(b) ishydrogen, alkyl, alkylalcohol, alkylcarboxyl, alkylcarboxylalkylester,acyl, acylalkyl, acylaryl, acyloxy, acyloxyalkyl, or acyloxyaryl; and Yis hydrogen or hydroxy; provided that R is other than allyl when R^(a)is methyl, R^(b) is propyl; and Y is hydrogen.
 9. The method of claim 8,wherein the subject is undergoing a hepatic resection or a livertransplantation.
 10. The method of claim 8, wherein R iscyclopropylmethyl, R^(a) is hydrogen, R^(b) is methyl, and Y is hydroxy.11. The method of claim 8, wherein R is methyl, R^(a) is methyl, R^(b)is methylacetylmethylester, and Y is hydrogen.
 12. The method of claim8, wherein R is allyl, R^(a) is methyl, R^(b) is ethylalcohol, and Y ishydrogen.